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Stabilized Polyunsaturated Compounds and Uses Thereof
BACKGROUND
Field
[0001] The present disclosure relates to the fields of biochemistry and
chemistry.
Some embodiments relate to stabilized polyunsaturated substances, composition
comprising such
stabilized polyunsaturated substances, and the therapeutic use thereof.
Description of the Related Art
[0002] in biological systems, the formation of potentially
physiologically-deleterious
reactive oxygen species (ROS) and reactive nitrogen species (RNS), may be
caused by a variety
of metabolic and/or environmental processes. By way of non-limiting example,
intracellular ROS
(e.g., hydrogen peroxide H202; superoxide anion 02-; hydroxyl radical OFF;
nitric oxide NO; and
the like) may be generated by several mechanisms: (i) by the activity of
radiation, both exciting
(e.g., UV-rays) and ionizing (e.g., X-rays); (ii) during xenobiotic and drug
metabolism; and (iii)
under relatively hypoxic, ischemic and catabolic metabolic conditions, as well
as by exposure to
hyperbaric oxygen. Protection against the harmful physiological activity of
ROS and RNS species
is mediated by a complex network of overlapping mechanisms and metabolic
pathways that utilize
a combination of small redox-active molecules and enzymes coupled with the
expenditure of
reducing equivalents.
[0003] Concentrations of ROS and RNS which cannot be adequately dealt
with by the
endogenous antioxidant system can lead to damage of lipids, proteins,
carbohydrates, and nucleic
acids. Changes in oxidative metabolism which lead to an increase in the
oxidizing environment
and the formation of potentially physiologically-deleterious ROS and RNS have
been generally
termed within the literature as "oxidative stress." There is a need for an
effective method to inhibit
or reduce such oxidative stress and further treatment of medical conditions
associated with
oxidative stress.
SUMMARY
[0004] Some embodiments relate to method of treating a subject having,
or at risk for,
a disease or condition associated with an impaired Phospholipase A2 Group VI
(PLA2G6)
activity, comprising administering to the subject an effective amount of a
substituted compound
selected from a polyunsaturated fatty acid, a polyunsaturated fatty acid
ester, a polyunsaturated
fatty acid thioester, a fatty acid amide, a polyunsaturated fatty acid
mimetic, a polyunsaturated
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fatty acid pro-drug, or combinations thereof, the substituted compound
comprises at least one
substitution that reduces oxidation of the substituted compound.
[0005] Some embodiments relate to method of treating a subject having,
or at risk for
an infantile neuroaxonal dystrophy (INAD) or PLA2G6 associated
neurodegeneration (PLAN),
comprising administering to the subject an effective amount of a substituted
compound selected
from a polyunsaturated fatty acid, a polyunsaturated fatty acid ester, a
polyunsaturated fatty acid
thioester, a fatty acid amide, a polyunsaturated fatty acid mimetic, a
polyunsaturated fatty acid
pro-drug, or combinations thereof, the substituted compound comprises at least
one substituent
that reduces oxidation of the substituted compound.
[0006] Some embodiments relate to method of treating a subject having,
or at risk for,
a disease or condition associated with a lysosomal storage disease (LSD),
comprising
administering to the subject an effective amount of a substituted compound
selected from a
polyunsaturated fatty acid, a polyunsaturated fatty acid ester, a
polyunsaturated fatty acid
thioester, a fatty acid amide, a polyunsaturated fatty acid mimetic, a
polyunsaturated fatty acid
pro-drug, or combinations thereof, the substituted compound comprising at
least one substituent
that reduces oxidation of the substituted compound.
[0007] Some embodiments relate to method of treating a subject having,
or at risk for
neuronal ceroid lipofuscinosis (NCL) type disease, comprising: administering
to the subject an
effective amount of a substituted compound selected from a polyunsaturated
fatty acid, a
polyunsaturated fatty acid ester, a polyunsaturated fatty acid thioester, a
fatty acid amide, a
polyunsaturated fatty acid mimetic, a polyunsaturated fatty acid pro-drug, or
a combination
thereof, the substituted compound comprising at least one substituent that
reduces oxidation of the
substituted compound.
100081 Some embodiments relate to method of treating a subject having,
or at risk for,
a sleeping disorder, comprising administering to the subject an effective
amount of a substituted
compound selected from a polyunsaturated fatty acid, a polyunsaturated fatty
acid ester, a
polyunsaturated fatty acid thioester, a fatty acid amide, a polyunsaturated
fatty acid mimetic, a
polyunsaturated fatty acid pro-drug, or combinations thereof, the substituted
compound
comprising at least one substituent that reduces oxidation of the substituted
compound.
BRIEF DESCRIPTION OF THE DRAWINGS
[0009] FIG. 1 summarizes the baseline and one year treatment status of a
patient
described in Example 1 (degree of impairment: (0) for severely impaired, (+1)
for moderately
impaired, and (+2) for mildly impaired or no impairment).
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DETAILED DESCRIPTION
[0010] Lipid peroxidation (LPO) is a self-propagating, free-radical
chain reaction that
amplifies toxic triggering effects in a variety of neurodegenerative
conditions. The present
disclosure relates to method of treating various diseases and conditions using
isotopically
modified polyunsaturated fatty acids or derivatives thereof, where these
isotopically modified
compounds have been stabilized via isotopic substitution at least one position
that reduces
oxidation of the compounds. These substituted compounds may be readily
incorporated into cell
membranes and may prevent, delays, or reverse lipid peroxidation and the
oxidative damage
caused by LPO.
Diseases associated with PLA2G6
[0011] The PLA2G6 gene encodes a group VIA calcium-independent
phospholipase
A2 beta enzyme that selectively hydrolyses glycerophospholipids to release
free fatty acids.
Mutations in PLA2G6 have been associated with disorders such as infantile
neuroaxonal
dystrophy (INAD), neurodegeneration with brain iron accumulation type II and
Karak syndrome.
PLA2G6 can be the causative gene in a subgroup of patients with autosomal
recessive early-onset
dystonia-palicinsonism. Neuropathological examination can show widespread Lewy
body
pathology and the accumulation of hypelphosphorylated tau.
[0012] In some studies, the mechanism of disease onset and progression
indicates that
lipid peroxidation pathways are the causes of disease phenotypes. In INAD
model, PLA2G6
mutations trigger accumulation of lipid peroxidation products of linoleic acid
and other PUFAs
leading to PLA2G6 associated neurodegeneration (PLAN). Studies using a fly
model of PLAN
have been conducted to address whether indeed toxic oxidized cardiolipin is
accumulating within
the mitochondrial membranes. The knock-out led to significant elevation in
lipid peroxidation
within the fly brain due to buildup of intracellular accumulations of stored
lipids. High
concentrations of lipid peroxidation products were observed.
[0013] INAD is a neurodegenerative disease with onset in infancy and
fatality in the
teenage years or in early adulthood. It is characterized neuropathologically
by axonal swelling and
the presence of spheroid bodies in the central and peripheral nervous systems
in addition to
hallmark cerebellar atrophy. Neurodegeneration with brain iron accumulation
comprises a
clinically and genetically heterogeneous group of disorders with a progressive
extrapyramidal
syndrome and high basal ganglia iron, and includes pantothenate kinase-
associated
neurodegeneration caused by mutations in PANK2 (neurodegeneration with brain
iron
accumulation type I). Post-mortem examination of the brain of a patient with
neurodegeneration
with brain iron accumulation associated with homozygous PLA2G6 mutations have
shown
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pathology with widespread Lewy bodies, dystrophic neurites and cortical
neuronal neurofibrillary
tangles.
[0014] Mutations in the PLA2G6 gene have been identified in most
individuals with
infantile neuroaxonal dystrophy. The PLA2G6 gene provides instructions for
making an enzyme
called an A2 phospholipase. This enzyme family is involved in metabolizing
phospholipids.
Phospholipid metabolism is important for many body processes, including
helping to keep the cell
membrane intact and functioning properly. The A2 phospholipase produced from
the PLA2G6
gene, sometimes called PLA2 group VI, helps to regulate the levels of a
compound called
phosphatidylcholine, which is abundant in the cell membrane. Mutations in the
PLA2G6 gene
impair the function of the PLA2 group VI enzyme. This impairment of enzyme
function may
disrupt cell membrane maintenance and contribute to the development of
spheroid bodies in the
nerve axons. Phospholipid metabolism problems have been seen in both this
disorder and a related
disorder called pantothenate kinase-associated neurodegeneration.
Lysosomal Storage Diseases
[0015] The term "lysosomal storage diseases" or "lysosomal storage
disorders" (LSD)
refers to a group of nearly fifty relatively rare inherited metabolic
disorders that result from defects
in lysosomal function as the result of deficiency of an enzyme, leading to the
inappropriate storage
of material in various cells of the body. These defects are related to
deficient cellular metabolism
of various types of lipids, glycoproteins and/or mucopolysaccharides. As a
result of LSD, excess
cell products that would ordinarily be broken down instead accumulate within
the cell to an
undesirable degree. Most lysosomal storage diseases are inherited in an
autosotnal recessive
manner. The symptoms of lysosomal storage disorders are generally progressive
over a period of
time. Some exemplary lysosomal storage diseases include: Gaucher disease
(Types I, II, and III),
Pompe disease (glycogen storage disease, including infantile form and a
delayed onset form),
GM2 gangliosidosis (including Tay-Sachs disease and Sandhoff disease), GM1
gangliosidosis,
and Niemann-Pick disease.
[0016] The term "neuronal ceroid lipofuscinosis" (NCL) is the general
name for a
family of genetically separate neurodegenerative lysosomal storage diseases
that result from
excessive accumulation of lipopigments (lipofuscin) in the body's tissues.
These lipopigments are
made up of fats and proteins. These lipofuscin materials build up in neuronal
cells and many
organs, including the liver, spleen, myocardium and kidneys.
[0017] GM1 gangliosidosis is a rare lysosomal storage disorder
characterized
biochemically by deficient beta-galactosidase activity and clinically by a
wide range of variable
neurovisceral, ophthalmological and dysmorphic features.
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100181 GM2 gangliosidoses are a group of three related genetic disorders
that result
from a deficiency of the enzyme beta-hexosaminidase. This enzyme catalyzes the
biodegradation
of fatty acid derivatives known as gangliosides. When beta-hexosaminidase is
no longer
functioning properly, the lipids accumulate in the nervous tissue of the brain
and cause problems.
GM2 gangliosidoses include Tay-Sachs disease, Sandhoff disease, and AB
variant.
100191 Tay-Sachs disease is a rare inherited disorder that progressively
destroys nerve
cells (neurons) in the brain and spinal cord. The most common form of Tay-
Sachs disease becomes
apparent in infancy. Other forms of Tay-Sachs disease are very rare. Signs and
symptoms can
appear in childhood, adolescence, or adulthood and are usually milder than
those seen with the
infantile form. Characteristic features include muscle weakness, loss of
muscle coordination
(ataxia) and other problems with movement, speech problems, and mental
illness. These signs and
symptoms vary widely among people with late-onset forms of Tay-Sachs disease.
Tay¨Sachs
disease is caused by a genetic mutation in the HEXA gene on chromosome 15. The
mutation
results in problems with an enzyme called beta-hexosaminidase A which results
in the buildup of
the molecule GM2 ganglioside within cells, leading to toxicity. Diagnosis is
by measuring the
blood hexosaminidase A level or genetic testing. In some embodiments of the
methods described
herein, selecting for treatment a subject having Tay-Sachs disease includes
measuring the blood
hexosaminidase A level, or testing genetic mutation in the HEXA gene.
100201 Sandhoff disease is a rare, autosomal recessive metabolic
disorder that causes
progressive destruction of nerve cells in the brain and spinal cord. The
disease results from
mutations on chromosome 5 in the HEXB gene, critical for the lysosomal enzymes
beta-N-
acetylhexosaminidase A and B. In some embodiments of the methods described
herein, selecting
for treatment a subject having Sandhoff disease includes testing genetic
mutation in the HEXB
gene.
100211 Gaucher's disease (GD) is a genetic disorder in which
glucocerebroside (a
sphingolipid, also known as glucosylceramide) accumulates in cells and certain
organs. The
disorder is characterized by bruising, fatigue, anemia, low blood platelet
count and enlargement
of the liver and spleen, and is caused by a hereditary deficiency of the
enzyme glucocerebrosidase
(also known as glucosylceramidase), which acts on glucocerebroside. When the
enzyme is
defective, glucocerebroside accumulates, particularly in white blood cells and
especially in
macrophages (mononuclear leukocytes). Glucocerebroside can collect in the
spleen, liver,
kidneys, lungs, brain, and bone marrow. This disease is caused by a recessive
mutation in the
GBA gene located on chromosome 1. Gaucher's disease is the most common of the
lysosomal
storage diseases. lit is a form of sphingolipidosis (a subgroup of lysosomal
storage diseases), as it
involves dysfunctional metabolism of sphingolipids. In some embodiments of the
methods
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described herein, selecting for treatment a subject having Gaucher's disease
includes testing
genetic mutation in the GBA gene.
[0022] Niemann-Pick disease is are a subgroup of lysosomal storage
disorders, which
is a group of inherited, severe metabolic disorders in which sphingomyelin
accumulates in
lysosomes in cells. Sphingomyelin is a component of cell membrane including
the organellar
membrane, so the enzyme deficiency blocks degradation of lipid, resulting in
the accumulation of
sphingomyelin within lysosomes in the macrophage-monocyte phagocyte lineage.
Mutations in
the SMPD1 gene cause Niemarm¨Pick disease types A and B. They produce a
deficiency in the
activity of the lysosomal enzyme acid sphingomyelinase, that breaks down the
lipid
sphingomyelin. Mutations in NPC1 or NPC2 cause Niemanir-Pick disease, type C
(NPC), which
affects a protein used to transport lipids. For type A and B, levels of
sphingomylinase can be
measured from a blood sample. To diagnose type C, a skin sample can help
determine whether
the transporter is affected.
[0023] Typical drugs target enzymes, proteins, or gene pathways.
However, many
biochemical processes are not controlled by enzymes. These processes are not
often addressed
therapeutically, in part, because modem drug discovery is usually based on
biochemical pathway
mapping informed by genomic analysis, and such approaches may be relatively
blind to non-
genetically encoded events. Non-enzymatic in vivo processes include a large
group of oxidation
reactions. The resulting oxidative damage is detrimental and, in diseased
cells, cannot be
controlled by antioxidants. Antioxidants are typically present in cells at
levels close to saturation
through enzymatically controlled active transport, and their concentrations
cannot be further
increased easily. In addition, excessive levels of antioxidants may interfere
with required redox
processes and result in a net detrimental effect. This may explain why
clinical trials of antioxidants
in humans often result in no positive or negative effects, even though the
disease aetiology is
oxidative in nature.
[0024] Lipid peroxidation may cause lysosomal instability and impairment
of
lysosome function, leading to LSD. As indicated above, LSD represents a class
of inborn
pathologies characterized by the accumulation of material in lysosomes. These
conditions can be
caused by the absence or reduced activity of lysosomal proteins, which results
in the lysosomal
accumulation of substances. Often, this material will be stored because
digestion is impaired due
to enzyme deficiency, but LSD can also arise when transport out of the
lysosomal compartment
is compromised. In some LSDs, the selection and transport of various
structurally damaged
moieties related to various lipid subclasses, for example sphingolipids, to
lysosomes for
processing can be compromised. Moreover, the accumulation of substances such
as highly
susceptible polyunsaturated fatty acids containing lipids can affect the
function of lysosomes and,
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further downstream, other organelles, resulting in secondary changes, such as
impairment of
autophagy, mitochondria] dysfunction, and inflammation. LSDs frequently
involve the central
nervous system, where neuronal dysfunction or loss results in mental
retardation, progressive
motor degeneration, and premature death.
[0025] The reactive oxygen species (ROS) play a pivotal role and are
perhaps common
mediators of cell death in many LSDs. Thus, up-regulation of apurinic
endonuclease 1 (APE1) (a
protein that repairs oxidative DNA damage) has been observed in Gaucher
fibroblasts (but not in
Gaucher bone marrow mesenchymal stromal cells). In the GM1 and GM2
gangliosidoses,
inducible nitric oxide synthase and nitrotyrosine are elevated in activated
microglia/macro-
phages, and ROS is elevated in Fabry disease models. Gene microarray analysis
from the
Niemann¨Pick disease type C 1 (NPC1) fibroblasts is consistent with enhanced
oxidative stress,
and elevated ROS and lipid peroxidation renders the fibroblasts more
susceptible to cell death
after an acute oxidative insult. In mucopolysaccharidosis type IIIB (MPSHIB),
enhanced oxidative
stress results in protein, lipid, and DNA oxidation, and an oxidative
imbalance is found in
mucopolysaccharidosis type I (MPS!). In neuronal ceroid lipofuscinose (NCL),
elevated ROS and
superoxide dismutase levels are suggested to be downstream to ER stress, a
significant increase
in manganese-dependent superoxide dismutase activity can be detected in
fibroblasts and brain
extracts from CLN6 sheep, and increased expression of 4-hydroxynonenal can be
detected in late
infant and juvenile forms of NCL.
[0026] The central role that oxidative stress plays in integrating other
cellular
pathways and stresses shows that it is most likely activated in LSDs as a
secondary biochemical
pathway, rather than as a direct result of accumulation of the primary
substrate. Moreover, the
possible role of oxidative stress may be of real significance in delineating
LSD pathology,
particularly as oxidative stress plays a central role in other better studied
neurodegenerative
conditions.
Sleeping Disorders
[0027] A very large proportion (around 40%) of the adult population is
affected by
various aspects of dyssomnia, either chronic or acute. Sleep plays an
important, multifunctional
role in physiological homeostasis. God N. Sleep Med Clin 2011;6:171. This
includes the clean-
up function, which sees to it that unwanted inter- and intra-neuronal
metabolic products
accumulated during the day are catabolized and removed. A significant fraction
of such debris
consists of oxidative stress generated materials such as various LPO
derivatives, on their own or
as conjugates with other biomolecules such a DNA, phospholipid head groups,
proteins or
peptides. Mathangi DC et al., Ann Neurosci 2012;19:161; Thamaraiselvi K et
al., Int. J. Biol. Med.
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Res. 2012;3:1754. It is well recognized that such derivatives are elevated,
and indeed can be used
as markers, of insufficient sleep. Weljie AM et al., PNAS USA 2015;112:2569.
[0028] External (lifestyle choices such as reduced sleep duration or
jetlag) or internal
(various sleeping disorders) factors adversely affect this elimination
process, resulting in non-
complete removal, or gradual accumulation, of LPO products, with ensuing
metabolic
pathologies, various neurological conditions, including, but not limited to,
psychosis and bipolar
disorder, as well as accelerated aging. Schmidt SM etal., Lancet Diabetes
EndocrinoL 2014;3:52.
A related problem is disturbance of circadian rhythmicity and oscillation,
which affects multiple
metabolic pathways. Moeller-Levet CS etal., PNAS USA 2013;110:E1132. This is
particularly
relevant to lipid processing, which is controlled to a very large degree by
circadian cycling. Chua
EC-P et aL, PNAS USA 2013;110:14468.
[0029] Elevated oxidative stress markers are associated with obstructive
sleep apnoea
syndrome, and in general with many other subclasses of dyssomnia. Passali D.
et al., Acta
OtorhinolcuyngoL Ital. 2015;35:420; Hachul DE etal., Climacteric 2006;9:312;
Gulec M etal.,
Prog Neuropsychopharmacol Biol Psychiatry 2012;37:247; Liang B et al., Eur Rev
Med
Pharmacol Sci 2013;17:2517; Semenova NV etal., Neuropsychiatry 2018;8:1452.
[0030] In some embodiments, the substituted compounds such as D-PUFAs
may be
used either alone or in combination with other treatments (including but not
limited to
antioxidants, melatonin, glycine, sleep medication, antidepressants, etc.) to
mitigate the side
effects of insufficient sleep and sleep disorders caused by various background
conditions,
including but not limited to, lifestyle related sleep deficiency; alcohol
related sleep deficiency;
idiopathic hypersomnia; narcolepsy; various sleep apneas; various parasomnias;
restless leg
syndrome; sleep state misperception; chronic fatigue syndrome (CFS) (also
referred to as myalgic
encephalomyelitis (ME)); mood disorders such as depression; anxiety disorders;
panic; psychoses
such as Schizophrenia; as well as circadian rhythm related sleep disorders,
including jetlag related
disorders and nightshift associated conditions. In some further embodiments,
the substituted
compounds may also help reducing the required amount of sleep and mitigate
somnolence. In
some further embodiments, the substituted compounds may also to useful to
improve, reduce, or
mitigate other physiological effects, side effects, or symptoms of a sleeping
disorder, such as
aching muscles; confusion; memory lapses or losses; depression; development of
false memory;
hypnagogic and hypnopompic hallucinations during falling asleep and waking;
hand tremor;
headaches; malaise; stye; pericnbital puffiness; increased blood pressure;
increased stress hormone
levels; increased risk of diabetes; lowering of immunity; increased
susceptibility to illness;
increased risk of fibromyalgia; irritability; rapid involuntary rhythmic eye
movement; obesity;
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seizures; temper tantrums in children; and symptoms similar to attention-
deficit hyperactivity
disorder and psychosis.
100311 Polyunsaturated lipids such as polyunsaturated fats, unlike
monounsaturated or
saturated fats, contain one or more bis-allylic positions ¨ that is -CH2
groups within the long
carbon chain of the fatty acid that are non-conjugated moieties between two
unsaturated double
bonds. These positions characterize PUFAs and are particularly susceptible to
oxidation stress by
hydrogen-abstraction to form a free radical. The radical, once formed, is much
more reactive than
the PUFA itself, and immediately reacts further, usually with oxygen, to form
peroxyl radicals,
and these are even better than the original disease trigger at propagating
more hydrogen-extraction
from l'UFAs (see Scheme 1).
100321 The chain reaction of PUFA autoxidation is illustrated in Scheme
1 as linoleic
acid chain reaction. More PUFAs are entering the cycle at the propagation
step. Abstraction of a
bis-allylic hydrogen is the rate-limiting step. Peroxides further degrade to
cytotoxic aldehydes that
further damage proteins and DNA, creating more peroxidation stress.
Scheme 1. Chemical Chain Reaction of Linoleic Acid Lipid Peroxidation
Initiation
(slow) if, OR
0
Oxygen
7" = ,r`oR
; , ===========================
..."^,..i ...,^-
...ejoFt
t
O'OsAs'
ti *NLiir flo
oft
1-4
=
o-oti
els
13 it) VR
Ternsnatkln
(recombinallon.
=RO antioxdsnte. etc)
i
100331 Instead of classical mechanisms of aberrant proteins, differing
expression
levels, or chemical toxicities can lead directly to disease damage to cells.
The factors such as
differing expression levels, chemical toxicities, and/or lipid peroxidation
may be triggers that do
not necessarily lead to clinical disease, unless and until amplified by a free
radical mechanism of
chemical oxidation of susceptible fatty acids. Since free radical lipid
oxidation mechanisms are
well understood, and known to involve a cycle of accelerating autocatalytic
damage, this common
mechanism, independent of the triggers used to initiate it, may be responsible
for the massive
amount of cellular damage across many indications.
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[0034] Down-regulating the free radical initiation of lipid peroxidation
(Scheme 1) can
both prevent and treat cellular damage responsible for disease, and even
reverse in vivo
phenotypes. Hence stabilizing lipids against such damage becomes a novel
treatment modality.
[0035] The mechanism of disease onset and progression indicates that
lipid
peroxidation pathways are linked to disease phenotypes. The pathway includes:
1) the high
concentration of ROS generated by cellular energy generation; 2) the
concentrated accumulation
of highly susceptible polyunsaturated fats in the lipid membranes; and 3) the
inadequate protection
by antioxidants due to various reasons including the hydrophobic nature of
membranes, which
limits antioxidant solubility and diffusion into the susceptible domains.
10036] The end metabolic products of damaged polyunsaturated fatty
acids, molecules
like 4-hydroxy-2- nonenal (4-HNE), 4-hydroxy hexenal (HHE), malondialdeyde
(MDA), and
many other reactive carbonyl compounds, are candidate markers of
neurodegeneration and
mitochondrial loss of function, and are observed in virtually all diseases in
which lipid
peroxidation is implicated.
Scheme 2. PUFA degradation products
HNE
OH
MIDA
0 õ 0
HHE 01-1
ITNE = 4-hydwxy-2- nonenai MDA = maiondialdeyde
HHE= 4-hydrk-my he.xelial
[0037] There is a broad body of literature supporting the role of lipid
peroxidation in
neurodegeneration with brain iron accumulation (NBIA) and related
neurodegenerative disease.
See Reed et al, "Lipid peroxidation and neurodegenerative disease," Free
Radical Biology &
Medicine 51(2011): 1302-1319. Recent literature also describes the role of
lipid peroxidation in
INAD, PKAN, and PLA2G6 disorders. See Kinghorn etal., "Loss of PLA2G6 leads to
elevated
mitochondrial lipid peroxidation and mitochondrial dysfunction," Brain
2015;138:1801-1816;
Kinghorn etal., "Mitochondrial dysfunction and defects in lipid homeostasis as
therapeutic targets
in neurodegeneration with brain iron accumulation," Rare Diseases 2016, VOL.
4, NO. 1,
el 128616. PLA2G6 has been implicated specifically in diseases with brain iron
accumulation,
such as Friedreich's ataxia, NBIA, and Alzheimer's disease¨to name a few¨are
even more
susceptible as iron is a Fenton reaction catalyst for the initiating event of
membrane lipid
peroxidation pathway.
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[0038] It is evident that lipid peroxidation plays a significant role in
LSD and/or NCL
and related neurodegenerative diseases. Malfunction in normal or oxidized
lipid processing
provokes LPO and exacerbates the toxicity of LPO products, imposing a systemic
toxic effect on
any lipid membrane containing structure, but particularly on PUFA rich
membranes. Examples of
LSD diseases include, but are not limited to, Sphingolipidoses, Ceramidase,
Father disease,
Krabbe disease (Infantile onset and Late onset), Galactosialidosis,
Gangliosides: gangliosidoses,
Alpha-galactosidase (including Fabry disease (alpha-galactosidase A),
Schindler disease (alpha-
galactosidase B)), Beta-galactosidase / GM1 gangliosidosis (Infantile,
Juvenile, and Adult /
chronic), GM2 gangliosidosis (AB variant, Activator deficiency, Sandhoff
disease (Infantile,
Juvenile, and Adult / chronic), Tay-Sachs (Juvenile hexosaminidase A
deficiency, Chronic
hexosaminidase A deficiency), Glucocerebroside (Gaucher disease, Type I, Type
II, Type III),
Sphingomyelinase (Lysosomal acid lipase deficiency, Early onset or Late
onset), Niemann-Pick
disease (Type A or Type B), Sulfatidosis, Metachromatic leukodystrophy,
Saposin B deficiency,
Multiple sulfatase deficiency, Mucopolysaccharidoses (Type I: MPS I Hurler
syndrome, MPS I S
Scheie syndrome, MPS I H-S Hurler-Scheie syndrome; Type II (Hunter syndrome);
Type III
(Sanfilippo syndrome) MPS III A (Type A), MPS III B (Type B), MPS III C (Type
C), MPS III
D (Type D); Type IV (Morquio): MPS WA (Type A), MPS NB (Type B); Type VI
(Maroteaux-
Lamy syndrome); Type VII (Sly syndrome); Type IX (hyaluronidase deficiency)),
Mucolipidosis
(Type I (sialidosis), Type II (I-cell disease), Type III (pseudo-Hurler
polydystrophy /
phosphotransferase deficiency), Type IV (mucolipidin 1 deficiency)), Lipidoses
(Niemann-Pick
disease, type C or Type D), Wo!man disease, Oligosaccharide (Alpha-
mannosidosis, Beta-
mannosidosis, Aspartylglucosaminuria, Fucosidosis), Lysosomal transport
diseases (Cystinosis,
Pycnodysostosis, Saila disease I sialic acid storage disease, Infantile free
sialic acid storage
disease), Glycogen storage diseases (Type II Pompe disease, Type Ilb Danon
disease), Cholesteryl
ester storage disease, and lysosomal disease. Examples of NCL type diseases
include but are not
limited to Type 1 Santavuori-Haltia disease / infantile NCL (CLN1 PPT1), Type
2 Jansky-
Bielschowsky disease / late infantile NCL (CLN2/LINCL TPP1), Type 3 Batten-
Spielmeyer-
Vogt disease / juvenile NCL (CLN3), Type 4 Kufs disease / adult NCL (CLN4),
Type 5 Finnish
Variant / late infantile (CLN5), Type 6 Late infantile variant (CLN6), Type 7
CLN7, Type 8
Northern epilepsy (CLN8), Type 8 Turkish late infantile (CLN8), Type 9
German/Serbian late
infantile (unknown), Type 10 Congenital cathepsin D deficiency (CTSD), and
Batten disease.
[0039] The lipid peroxidation chain reaction is the target of the
substituted compounds
described herein. This chain reaction results in cell damage, death and
disease. To halt this damage
process substituted compounds as described herein can target the root cause of
disease, the
amplification of the original disease trigger by lipid peroxidation. Since
PUFAs also turn over in
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diseased as well as normal cells, substituted compounds as described herein
can both maintain
and restore health and function to them.
100401 The initiation event of the lipid peroxidation chain reaction is
caused by ROS
abstracting a hydrogen off a bis-allylic (between the double bonds) methylene
carbon in the lipid-
this is the rate determining step of the chain reaction in lipid peroxidation.
If one could slow down
the initiation rate, it would have a large effect down-regulating PUFA
oxidation by eliminating all
of the downstream multiplying 'cycles' of damage from each abstraction.
100411 The initial abstraction rate can be reduced by replacing hydrogen
atoms at bis-
allylic methylene sites with deuterium atoms. Deuterium is naturally present
and is recognized by
living systems as a normal variation of hydrogen (typically hydrogen in all
natural substances
consists of 1 deuterium per 7000 hydrogens). Deuterium is also responsible for
a well-known
"isotope effect" (1E): reactions involving cleavage of a C-H bond are slowed
down substantially
when H is replaced with D. This substitution reduces the ability of the C-H
bond to be broken.
100421 In some embodiments, substituted compounds as described herein
(e.g.,
PUFAs) that are specifically substituted with deuterium at bis-allylic
positions can be made in
large quantities using well-optimized drug synthetic methods. This
modification is both "natural"
(deuterium exists in nature) and "game-changing": whereas the lipid
peroxidation process is
autocatalytic, the stabilization of the initiating step is 'anti-' catalytic,
causing at each step a
multiplicative > 10-fold isotope reduction, essentially shutting down the
chain process quickly.
Hence, the susceptible target bonds of the chain reaction are "fire-proofed"
against the damage of
ROS. Importantly, enzymatic processes involving PUFAs, such as I3-oxidation,
transformations
involving other enzymes (all stoichiometric 1:1 enzyme: substrate reactions)
are largely
unaffected. In addition, this 'fireproofing' process requires only a fraction
of the total PUFAs be
deuterated for the chain reaction of lipid peroxidation to be effectively shut
down.
100431 Substituted compounds as described herein that are deuterated
(e.g., deuterated
PUFAs) thus represent a novel type of sensitive and specific drug which is
structurally similar to
corresponding compounds having only a natural level of deuteration, but they
prevent damaging,
non-enzymatic oxidation processes without interfering substantially with
biologically necessary
enzymatic transformations. For example, because PUFAs in membranes turn over
rapidly¨even
when the cells do not¨deuterated PUFAs rapidly replace the original hydrogen
containing
molecules in all compartments in all tissues. All of the active transport used
to transfer normal
PUFAs from orally ingested foods work the same on deuterated PUFAs, and
transport them
wherever they are needed. As a result, D-PUFAs rapidly incorporate into brain,
retina, and other
difficult to treat tissues.
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10044] The substituted compounds as described herein (e.g., deuterated
PUFAs,
11,11-D2-linoleic acid ethyl ester) is unique in drug discovery and treatment.
Some PUFAs, such
as linoleic acid, are part of the human diet that have no pharmacological
effect, yet in the
deuterated form they may act as sensitive and specific drugs. These type of
substituted compounds
also do not have any observable side effects.
10045] Linoleic acid (LA) is essential for human diets and is designated
as GRAS
without a known toxic upper limit for nutritional use. LA was identified in
the 1920s and there
have been more than 1,300 published human studies of LA. There have been more
than 23,000
published human studies of omega-3 PUFAs of which about 2,500 were randomized
controlled
clinical trials comparing omega-3 PUFAs to LA. No LA-related safety issues
were identified in
these studies.
[0046] Substituted compounds as described herein can be effective in
treating a
disease or condition associated with a lysosomal storage disease or a
condition associated with
impaired PLA2G6 activity. The effect of the substituted compounds (e.g.,
deuterated LA and ester
thereof) in promoting cell survival, reducing and/or preventing the damage
arising from the free
radical chain reaction has been demonstrated. Coenzyme Q deficient coq mutant
yeast strains are
highly sensitive to oxidation damage from exogenous PUFAs because they lack
antioxidant
control and the critical hydrophobic intracellular mitochondria' membrane
domains are not
accessible to other hydrophilic antioxidants. In a coq mutant yeast model,
cells that are otherwise
healthy when grown on MUFAs (mono-unsaturated fatty acids) and SFAs (saturated
fatty acids),
do not grow well in the presence of a PUFA (e.g. LA). However, a substituted
compound (e.g.,
deuterated LA and ester thereof) is effective in increasing and/or preserving
the viability of the
cells.
[0047] In addition, a substituted compound as described herein can be
effective in
reducing and preventing oxidative stress and damages associated with iron
accumulation. Yeast,
murine, and human in vitro models of Friedreich's ataxia (FRDA) demonstrate
that a substituted
compound as described herein (e.g., D-PUFA and ester thereof) has been
effective in managing
the oxidative stress associated with increased iron. In FRDA damaging iron
accumulations are
observed in the brain tissue, similar to that of INAD. In the yeast model of
FRDA, treatment with
a substituted compound as described herein (e.g., D-PUFA and ester thereof)
and D4-ALA
(deuterated linolenic acid such as 11,11,14,14-D4-linolenic acid) can result
in decreased lipid
peroxidation. In the murine FRDA cell model, a substituted compound as
described herein (e.g.,
11,11-D2-linoleic acid, 11,11,14,14-D4-linolenic acid, or ester thereof) can
protect cells from loss
of viability. In the human fibroblast FRDA model, a substituted compound as
described herein
(e.g., 11,11-D2-linoleic acid, 11,11,14,14-D4-linolenic acid, or ester
thereof) can rescue cells from
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loss of viability. See Cotticelli et al., Redox Biology. 2013 (1):398-404. In
another murine model
of FRDA, treatment with D4-ALA can prevent lipid peroxidation. See Abeti et
al., Cell Death
Di s. 2016 May 26; 7: e2237.
[0048] The effect of substituted compounds as described herein (e.g., D-
PUFA and
ester thereof) on mitochondria' function wider oxidative stress conditions was
reported by
Andreyev. See Andreyev et al., Free Radic Biol Med. 2015 Jan 8. S0891-
5849(15)00003-9. Levels
of F2-IsoProstane oxidation products can be decreased dramatically in the
deuterated-treated cells,
indicating that treatment with the substituted compounds as described herein
can decrease the free
radical chain processes in a cell as a whole. In H9C2 myoblasts, treatment
with t-ButOOH can
cause both respiratory inhibition and increased membrane leak. Substituted
compounds as
described herein (e.g., D-PUFA and ester thereof) can protect mitochondria'
function from stress
caused by t-ButO0H; maximal respiration is preserved and/or increase in
membrane leak is
diminished. Other oxidative stress paradigms can also be operative and D4-ALA
and ester thereof
can be protective against oxidative stress, confirming that the combination of
non-deuterated and
deuterated PUFAs can be effective in protecting against oxidative stress. A
small amount of D-
PUFA can provide significant protection against a very severe oxidative stress
induced by Fe2+ in
the presence of an unprotected polyunsaturated fatty acid.
[0049] An in vivo model of mitochondria' dysfunction shows that
substituted
compounds as described herein (e.g., D-PUFA and ester thereof) reduce
oxidative stress-related
injury. In a model of Parkinson's disease, C57BL/6 mice are treated with the
neurotoxin 1-methyl-
4-pheny1-1,2,3,6-tetrahydropyridine (MPTP). See Shchepinov et al., Toxicol
Lett. 2011 Nov
30;207(2):97-103. MPTP's active metabolite, MPP+, inhibits complex I of the
mitochondria'
electron transport chain. Measurement of brain deuterium concentration after
administration of
substituted compounds as described herein (e.g., D-PUFA and ester thereof) in
the diet for 12-13
days indicates that deuterium is incorporated. The animals fed with
substituted compounds as
described herein (e.g., D-PUFA and ester thereof) show less MPTP effect when
compared with
the controls. Treatment with substituted compounds as described herein (e.g.,
D-PUFA and ester
thereof) may also rescue the dopaminergic phenotype.
[0050] Single- and repeat-dose studies are conducted in mice and rats
using both oral
gavage and dietary administration of substituted compounds as described herein
(e.g., D-PUFA
and ester thereof) for up to 26 weeks. The substituted compounds studied
(e.g., 11,11-D2-linoleic
acid) were well tolerated in all of the studies conducted. The NOAELs
established in the 8- and
26-week studies correspond to an average consumption of the substituted
compounds studied in
the amount of -362 and -452 mg/kg, respectively. The high dose diets in these
studies contained
no natural LA. There were no signs observed in this study of essential fatty
acid (linoleic acid)
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deficiency, which is characterized by changes, notably to the skin including
alopecia and scaly
tails, which appear within months of sustained feeding of a diet lacking LA.
This is consistent
with the substituted compounds as described herein (e.g., D-PUFA and ester
thereof) (di-deutero
linoleic acid) being, for dietary purposes, equivalent to and biologically
interchangeable with
normal dietary LA as a sole source of LA by the rats in this study. Analysis
of tissue uptake and
distribution indicates that administration of substituted compounds as
described herein (e.g., D-
PUFA and ester thereof) do not appear to alter the enzymatic processing or
availability of PUFAs
as measured in the heart, brain, lung, kidney, and liver. Substituted
compounds as described herein
(e.g., D-PUFA and ester thereof) and their derivatives are incorporated into
tissues and do not
result in any significant morphological or functional changes in the treated
animals versus their
controls. The data show that the enzymatic processing of substituted compounds
as described
herein (e.g., D-PUFA and ester thereof), and the subsequent selective PUFA
species incorporation
patterns for each of the tissues tested, are the same for the low dose, high
dose and control groups
for both 8-week and 26-week studies. The body weights, organ weights, percent
of each organ
composed of PUFAs, distribution of PUFA species in each tissue, and the PUFA
composition of
red blood cells are not changed versus controls.
100511 These data indicate that substituted compounds as described
herein (e.g., D-
PUFA and ester thereof) can be effective in inhibiting the free radical
degradation of lipids but do
not impact the metabolic enzymatic processing of lipids. Thus, substituted
compounds as
described herein (e.g., D-PUFA and ester thereof) appear to be processed
identically to dietary-
sourced essential fatty acid LA, and hence share the well-known
characteristics and safety profile
of LA.
100521 Substituted compounds as described herein (e.g., D-PUFA and
esters thereof)
have been demonstrated in many neurodegenerative disease preclinical models to
mitigate both
cell death and disease symptoms. The gene defect underlying >90% of INAD
disease, PLA2G6,
causes increased cell death due from inability to mop up lipid peroxidation.
These effects can be
reversed over controls in INAD stem cell and fibroblast studies with D-PUFA
dosing, and
climbing performance in a drosophila model improved. Since D-PUFAs have been
proven safe in
preclinical toxicity studies and a clinical phase 1/2 study in Friedreich's
ataxia, and showed benefit
in multiple INAD models, the treatment of a disease or condition associated
with an impaired
Phospholipase A2 Group VI (PLA2G6) activity such as INAD and PLAN, or a
disease or
condition associated with an-lysosomal storage disease and/or NCL disease can
also be effective.
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Definition
[0053] The section headings used herein are for organizational purposes
only and are
not to be construed as limiting the subject matter described.
[0054] Unless defined otherwise, all technical and scientific terms used
herein have
the same meaning as is commonly understood by one of ordinary skill in the
art. The use of the
term "including" as well as other forms, such as "include", "includes," and
"included," is not
limiting. The use of the term "having" as well as other forms, such as "have",
"has," and "had,"
is not limiting. As used in this specification, whether in a transitional
phrase or in the body of the
claim, the terms "comprise(s)" and "comprising" are to be interpreted as
having an open-ended
meaning. That is, the above terms are to be interpreted synonymously with the
phrases "having
at least" or "including at least." For example, when used in the context of a
process, the term
"comprising" means that the process includes at least the recited steps, but
may include additional
steps. When used in the context of a compound, composition, formulation, or
device, the term
"comprising" means that the compound, composition, formulation, or device
includes at least the
recited features or components, but may also include additional features or
components.
[0055] As used herein, common abbreviations are defined as follows:
ANOVA analysis of variance
BID Twice daily
WAD Infantile neuroaxonal dystrophy
LOTS Late onset Tay-Sachs disease
LPO Lipid Peroxidation
LSD Lysosomal storage disease or disorder
RBC Red blood cell
PLA2G6 Phospholipase A2 Group VI
PLAN PLA2G6 associated neurodegeneration
PK Pharmacokinetics
PUFA Polyunsaturated Fatty Acid
T25FW Timed 25-foot walk
[0056] The term "about" as used herein, refers to a quantity, value,
number,
percentage, amount, or weight that varies from the reference quantity, value,
number, percentage,
amount, or weight by a variance considered acceptable by one of ordinary skill
in the art for that
type of quantity, value, number, percentage, amount, or weight. In various
embodiments, the
term "about" refers to a variance of 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%,
3%, 2% or 1%
relative to the reference quantity, value, number, percentage, amount, or
weight.
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100571 The term "oral dosage form," as used herein, has its ordinary
meaning as
understood by those skilled in the art and thus includes, by way of non-
limiting example, a
formulation of a drug or drugs in a form administrable to a human, including
pills, tablets, cores,
capsules, caplets, loose powder, solutions, and suspensions.
[0058] As used herein, the term "ester" refers to the structure
¨C(=0)0R, wherein R
may include unsubstituted or substituted C1-30 alkyl (branched or straight),
unsubstituted or
substituted substituted C6-10 aryl, unsubstituted or substituted 5 to 10
membered heteroaryl,
unsubstituted or substituted C3-10 carbocyclyl, or unsubstituted or
substituted 3 to 10 membered
heterocyclyl.
[0059] As used herein, the term "thioester" refers to the structure
¨C(=0)SR, wherein
R may include unsubstituted or substituted C1-30 alkyl (branched or straight),
unsubstituted or
substituted substituted C6-10 aryl, unsubstituted or substituted 5 to 10
membered heteroaryl,
unsubstituted or substituted C3-10 carbocyclyl, or unsubstituted or
substituted 3 to 10 membered
heterocyclyl.
[0060] As used herein, the term "amide" refers to the structure -
C(0)NRIR2 or
¨S(0)NRIR2, and RI and R2 can independently be unsubstituted or substituted C1-
30 alkyl
(branched or straight), unsubstituted or substituted substituted C6-10 aryl,
unsubstituted or
substituted 5 to 10 membered heteroaryl, unsubstituted or substituted C3-10
carbocyclyl, or
unsubstituted or substituted 3 to 10 membered heterocyclyl.
100611 "Subject" as used herein, means a human or a non-human mammal,
e.g., a dog,
a cat, a mouse, a rat, a cow, a sheep, a pig, a goat, a non-human primate or a
bird, e.g., a chicken,
as well as any other vertebrate or invertebrate.
[0062] The term "pediatric patient" as used herein means a human patient
that is 17
years old or younger. In certain non-limiting embodiments, the patient is 16
years old or younger,
or 15 years old or younger, or 14 years old or younger, or 13 years old or
younger, or 12 years old
or younger, or 11 years old or younger, or 10 years old or younger, or 9 years
old or younger, or
8 years old or younger, or 7 years old or younger, or 6 years old or younger,
or 5 years old or
younger, or 4 years old or younger, or 3 years old or younger, or 2 years old
or younger, or 1 year
old or younger, or 6 months old or younger, or 4 months old or younger, or 2
months old or
younger, or 1 months old or younger. In particular embodiments, the pediatric
patient is between
about 12 to about 17 years of age. In one embodiment, the pediatric patient
has an age selected
from the group consisting of between about 12 to about 17 years of age and
about 2 years of age
or younger.
100631 As used herein, the act of "providing" includes supplying,
acquiring, or
administering (including self-administering) a composition described herein.
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[0064] As used herein, the term "administering" a drug includes an
individual
obtaining and taking a drug on their own. For example, in some embodiments, an
individual
obtains a drug from a pharmacy and self-administers the drug in accordance
with the methods
provided herein.
[0065] The term "therapeutically effective amount" as used herein,
refers to an amount
of a substituted compound described herein sufficient to treat, ameliorate a
disease or condition
described herein, or to exhibit a detectable therapeutic effect. The effect
may be detected by any
means known in the art. In some embodiments, the precise effective amount for
a subject can
depend upon the subject's body weight, size, and health; the nature and extent
of the condition;
and the therapeutic or combination of therapeutics selected for
administration. Therapeutically
effective amounts for a given situation may be determined by routine
experimentation that is
within the skill and judgment of the clinician. In some embodiments, the
substituted compound is
a polyunsaturated acid (PUFA) or an ester, thioester, amide, or other prodrug
thereof, or
combinations thereof for treating, or ameliorating the diseases or conditions
described herein. In
some further embodiment, the substituted compound is 11,11-D2-linoleic acid or
an ester thereof.
[0066] "Treat," "treatment," or "treating," as used herein refers to
administering a
compound or pharmaceutical composition to a subject for prophylactic and/or
therapeutic
purposes. The term "prophylactic treatment" refers to treating a subject who
does not yet exhibit
symptoms of a disease or condition, but who is susceptible to, or otherwise at
risk of, a particular
disease or condition, whereby the treatment reduces the likelihood that the
patient will develop
the disease or condition. The term "therapeutic treatment" refers to
administering treatment to a
subject already suffering from a disease or condition.
[0067] The pharmaceutical composition described herein are preferably
provided in
unit dosage form. As used herein, a "unit dosage form" is a
composition/formulation containing
an amount of a compound that is suitable for administration to an animal,
preferably mammal
subject, in a single administration, according to good medical practice. The
preparation of a single
or unit dosage form however, does not imply that the dosage form is
administered once per day
or once per course of therapy, or that the unit dosage form contains all of
the dose to be
administered at a single time. Such dosage forms are contemplated to be
administered once, twice,
thrice or more per day, and may be given more than once during a course of
therapy, though a
single administration is not specifically excluded. In addition, multiple unit
dosage forms may be
administered at substantially the same time to achieve the full dose intended
(e.g., two or more
tablets may be swallowed by the patient to achieve a complete dose). The
skilled artisan will
recognize that the formulation does not specifically contemplate the entire
course of therapy and
such decisions are left for those skilled in the art of treatment rather than
formulation.
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[0068] In any of the embodiments described herein, methods of treatment
can
alternatively entail use claims, such as Swiss-type use claims. For example, a
method of treating
a subject having an impaired PLA2G6 activity can alternatively entail the use
of a compound in
the manufacture of a medicament for the treatment of the disease(s) or
condition(s) described
herein, or a compound for use in the treatment of the disease(s) or
condition(s) described herein.
Methods of Treatment
[0069] Some embodiments relate to a method of treating a subject having,
or at risk
for, a disease or condition associated with an impaired Phospholipase A2 Group
VI activity,
comprising: selecting a subject having, or at risk for, a disease or condition
associated with an
impaired Phospholipase A2 Group VI activity; and administering to the subject
an effective
amount of a substituted compound selected from the group consisting of a
polyunsaturated fatty
acid, a polyunsaturated fatty acid ester, a polyunsaturated fatty acid
thioester, a fatty acid amide,
a polyunsaturated fatty acid mimetic, a polyunsaturated fatty acid pro-drug,
and combinations
thereof, wherein the substituted compound comprises at least one substituent
that reduces
oxidation of the substituted compound. In some embodiments, the subject has
infantile
neuroaxonal dystrophy or PLA2G6 associated neurodegeneration. In one
embodiment, the subject
has infantile neuroaxonal dystrophy. In some such embodiments, the infantile
neuroaxonal
dystrophy is caused by PLA2G6 mutation.
[0070] Some embodiments relate to a method of treating a subject having,
or at risk
for, a disease or condition associated with a lysosomal storage disease and/or
neuronal ceroid
I ipofuscinosis disease, comprising: selecting a subject having, or at risk
for, a disease or condition
associated with a lysosomal storage disease or neuronal ceroid lipofuscinosis;
and administering
to the subject an effective amount of a substituted compound selected from a
polyunsaturated fatty
acid, a polyunsaturated fatty acid ester, a polyunsaturated fatty acid
thioester, a fatty acid amide,
a polyunsaturated fatty acid mimetic, a polyunsaturated fatty acid pro-drug,
or combinations
thereof, wherein the substituted compound comprises at least one substituent
that reduces
oxidation of the substituted compound. In some embodiments, the subject has
Tay-Sachs, Gaucher
disease, Sandhoff disease, or Niemarm-Pick disease. In one embodiment, the
subject has Tay-
Sachs disease, for example, late onset Tay-Sachs disease. In some such
embodiments, Tay-Sachs
disease is caused by genetic mutation in the HEXA gene. In some embodiments,
the LSD is GM!
gangliosidosis. In some embodiments, the LSD is GM2 gangliosidosis. In some
embodiments, the
LSD is sphingolipidose disease.
[0071] Some embodiments relate to a method of treating a subject having,
or at risk
for, a sleeping disorder, comprising: selecting a subject having, or at risk
for, a sleeping disorder;
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and administering to the subject an effective amount of a substituted compound
selected from the
group consisting of a polyunsaturated fatty acid, a polyunsaturated fatty acid
ester, a
polyunsaturated fatty acid thioester, a fatty acid amide, a polyunsaturated
fatty acid mimetic, a
polyunsaturated fatty acid pro-drug, and combinations thereof, wherein the
substituted compound
comprises at least one substituent that reduces oxidation of the substituted
compound. In some
embodiments, the subject has acute or chronic dyssomnia. In some embodiments,
the subject has
obstructive sleep apnoea syndrome.
[0072] In some embodiments of the methods described herein, the
administering step
comprises repeated administration. In some embodiments, the subject has or is
at risk for at least
one of neuropathy or a neurodegenerative disease and the amount of the
substituted compound is
effective to prevent, ameliorate or inhibit the progression of neuropathy or
the neurodegen.erative
disease.
[0073] In some embodiments of the methods described herein, the
substituted
compound comprises one or more isotopes, and the amount of the isotope is
significantly above
the naturally-occurring abundance level of the isotope. For example, in some
embodiments, the
amount of the isotope is two or more times greater than the naturally-
occurring abundance level
of the isotope. In some embodiments, the isotope is selected from deuterium,
DC, and a
combination thereof. In some embodiments, the isotope atom is deuterium. The
substituted
compound, for example, isotopically modified PUFAs such as deuterated PUFAs
may reduce
oxidation by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%,
60%, 65%,
70%, 75%, 80%, 85%, 90% or 95%.
[0074] In some embodiments, the method described herein comprises
identifying or
selecting from treatment a subject having an impaired PLA2G6 activity. In some
such
embodiments, identifying or selecting the subject having the impaired PLA2G6
activity may
include sequencing the subject's DNA or using a genetic test to identify and
screen for patients
having a mutation of PLA2G6 gene. In some embodiments, identifying a patient
having an
impaired PLA2G6 activity is established in a proband by identification of
biallelic pathogenic
variants in PLA2G6 on molecular genetic testing. In some embodiments,
identifying a patient
having an impaired PLA2G6 activity can be established in a proband with no
identified PLA2G6
pathogenic variants by electron microscopic examination of nerve biopsies for
dystrophic axons
(axonal spheroids).
[0075] In some embodiments, the method described herein comprises
identifying or
selecting for treatment a subject having LSD and/or NCL. In some such
embodiments, identifying
or selecting the subject having LSD and/or NCL may include sequencing the
subject's DNA or
using a genetic test to identify and screen for patients having a gene
mutation associated with a
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LSD and/or NCL type disease. In some such embodiments, identifying or
selecting the subject
having LSD and/or NCL comprises sequencing the subject's DNA or using a
genetic test to
identify and determine the expression or activity of PGRN or detecting one or
more mutation in
the genomic DNA or gene encoding PGRN. In some embodiments, identifying or
selecting the
subject having LSD and/or NCL comprises sequencing the subject's DNA or using
a genetic test
to identify and determine the expression or activity of PGRN or detecting one
or more mutation
in the genomic DNA or gene encoding HEX (e.g., HEXA, HEXB, or HEXS). In some
further
embodiments, identifying or selecting the subject having LSD and/or NCL
comprises sequencing
the subject's DNA and detecting one or more mutation in the genomic DNA or
gene encoding
HEX (e.g., HEXA, HEXB, or HEXS). In some embodiments, the gene mutation
associated with
a LSD and/or NCL can be GBA mutation. In some embodiments, the gene mutation
associated
with a LSD and/or NCL can be PGRN mutation. In some embodiments, identifying
the subject
having LSD and/or NCL comprises sequencing the subject's DNA or using a
genetic test to
identify and determine the expression or activity of SM PD1, NPC1, or NPC2 or
detecting one or
more mutation in the genomic DNA or gene encoding SMPD1, NPC1, or NPC2.
[0076] In some embodiments of the methods described herein, the subject
has or is at
risk for at least one of a neuropathy or a neurodegenerative disease
associated with the impaired
PLA2G6 activity. In some embodiments, the subject has an infantile neuroaxonal
dystrophy
(INAD) or PLA2G6 associated neurodegeneration (PLAN). In some embodiments, the
neuropathy or a neurodegenerative disease associated with the impaired PLA2G6
activity does
not include Alzheimer's disease. In some embodiments, neuropathy or a
neurodegenerative
disease associated with the impaired PLA2G6 activity does not include
Parkinson's disease.
100771 In some embodiments, the amount of the substituted compound
administered
to the subject is effective to alleviate one or more symptoms of the disease
or condition associated
with the impaired Phospholipase A2 Group VI (PLA2G6) activity. In some such
embodiments,
the symptom of the disease or condition is selected from the group consisting
of hypotonia,
nystagmus, strabismus, psychomotor regression, and low spontaneous motor
activity.
[0078] In some embodiments, the amount of the substituted compound
administered
to the subject is effective to alleviate one or more symptoms associated with
the LSD and/or NCL.
The symptoms for LSD and/or NCL may be different depending on the patient
conditions. In some
embodiments, the symptom of the disease or condition is at least one selected
from the group
consisting of difficulties with physical movement (e.g., joint stiffness and
pain), seizures,
dementia, mental retardation, high fatality, problems vision (e.g., blindness)
or hearing (deafness),
and problem with bulbar function.
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[0079] In some embodiments, the amount of the substituted compound
administered
to the subject is effective to alleviate one or more symptoms or side effects
of a sleeping disorder
or insufficient sleep. In some such embodiments, the side effects or symptom
of a sleeping
disorder or insufficient sleep are selected from the group consisting of
aching muscles; confusion;
memory lapses or losses; depression; development of false memory; hypnagogic
and
hypnopompic hallucinations during falling asleep and waking; hand tremor;
headaches; malaise;
stye; perioibital puffiness; increased blood pressure; increased stress
hormone levels; increased
risk of diabetes; lowering of immunity; increased susceptibility to illness;
increased risk of
fibromyalgia; irritability; rapid involuntary rhythmic eye movement; obesity;
seizures; temper
tantrums in children; and symptoms similar to attention-deficit hyperactivity
disorder and
psychosis.
[0080] In some further embodiments, the amount of the substituted
compound
administered to the subject is effective to increase the muscle functions of
the subject. In some
embodiments, the muscle function is selected from the group consisting of eye
tracking, control,
lifting, fine motor skill, and muscle strength.
[0081] In some further embodiments, the amount of the substituted
compound
administered to the subject is effective to increase the neural function of
the subject. In some
embodiments, the neural function is selected from the group consisting of
responsiveness to verbal
commands, bulbar function, and verbal cognition. In some embodiments, the
neural function is
the bulbar function.
[0082] In some embodiments, administration of the substituted compounds
as
described herein can be used in combination with one or more additional
therapies for treating
INAD selected from a pharmacologic treatment of spasticity and seizures; a
trial of oral or
intrathecal baclofen for dystonia associated with atypical INAD; a treatment
by a psychiatrist for
those with later-onset neuropsychiatric symptoms; a fiber supplement and/or
stool softener
treatment for constipation; control of secretions with transdermal scopolamine
patch as needed;
feeding modifications as needed to prevent aspiration pneumonia and achieve
adequate nutrition,
and a combination thereof. In some embodiments, administration of a
substituted compound as
described herein can be used in combination with one or more additional
therapies for treating
PLAN selected from the group consisting of treatment with dopaminergic agents;
treatment of
neuropsychiatric symptoms by a psychiatrist; evaluation by physical therapy
for management of
postural instability and gait difficulties; occupational therapy to assist
with activities of daily
living; feeding modifications as needed to prevent aspiration pneumonia and
achieve adequate
nutrition, and a combination thereof.
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10083] In some embodiments, administration of the substituted compounds
as
described herein can be used in combination with one or more additional
therapies for treating
LSD and/or NCL diseases. For example, a subject suffering from a sleeping
disorder described
herein may be administered with antioxidants, melatonin, glycine, sleep
medication,
antidepressant to improve or regulate sleep-wake cycle and/or mitigate the
side effects associated
with a sleeping disorder or insufficient sleep.
Doses
10084] In some embodiments of the methods described herein, the
therapeutically
effective amount of a substituted compound administered to the subject is
about 0.1g, 0.2g, 0.5g,
1.0g, 1.5g, 2.0g, 2.5g, 3.0g, 3.5g, 4.0g, 4.5g, 5.0g, 5.5g, 6.0g, 6.5g, 7.0g,
7.5g, 8.0g, 8.5g, 9.0g,
9.5g, 10g, 10.5g, 11g, 11.5g, 12g, 12.5g, 13g, 13.5g, 14g, 14.5g, 15g, 15.5g,
16g, 16.5g, 17g,
17.5g 18g, 18.5g, 19g, 19.5g, or 20g, or a range defined by any of the two
preceding values. In
some embodiments, the amount of the substituted compound administered is from
about 0.1g to
about 20g, from about 1 g to about 10g, from 2g to about 5g. In some further
embodiments, the
amount of the substituted compound administered is from about 1.8g to about
4.5g. [n some
embodiments, the substituted compound is in a single unit dosage form. In some
other
embodiments, the substituted compound is in two or more unit dosage forms
(i.e., a divided dose).
For example, where a dose is about 5g, it may be provided in the form of four
or five tablets, each
containing about 1.25g or lg of the substituted compound. In some such
embodiments, a dose of
lg to lOg comprises administering 1, 2, 3, 4 or 5 unit dosage forms each
comprising from about
lg to about 2g of the substituted compound, or about 2,3, or 4 unit dosage
forms each comprising
from about 0.5g to about 2.5g of the substituted compound. In another example,
a dose of 2g to
5g comprises administering 1, 2, 3, 4 or 5 unit dosage forms each comprising
from about 1g to
about 2g of the substituted compound. In some embodiments, the unit dosage
form is a tablet, a
capsule, a pill, or pellets. In some further embodiment, the unit dosage form
for oral
administration, i.e., oral dosage form.
10085] In some embodiments of the methods described herein, the
substituted
compound may be administered once per day. In some other embodiments, the
substituted
compound may be administered two or more times per day, for example, twice a
day or three
times a day. In some embodiments, the therapeutically effective amount of the
substituted
compound administered per day is about 1.0g, 2.0g, 3.0g, 3.5g, 4.0g, 4.5g,
5.0g, 5.5g, 6.0g, 6.5g,
7.0g, 7.5g, 8.0g, 8.5g, 9.0g, 9.5g, 10g, 10.5g, 11g, 11.5g, 12g, 12.5g, 13g,
13.5g, 14g, 14.5g, 15g,
15.5g, 16g, 16.5g, 17g, 17.5g 18g, 18.5g, 19g, 19.5g, 20g, 25g, 30g, 35g, 40g,
45g, or 50g, or a
range defined by any of the two preceding values. In some such embodiments,
the amount of the
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substituted compound administered per day is from about lg to about 20g, from
about 2g to about
10g, from about 3g to about 8g, from about 4g to about 7g, or from about 5g to
about 6g. In one
embodiment, the amount of 11,11-D2-linoleic acid or the ester thereof
administered per day is
from about 2g to about 10g. In another embodiment, the amount of 11,11-D2-
finoleic acid or the
ester thereof administered per day is from about 1.8 g to about 9g.
100861 In some embodiments of the method described herein, the
substituted
compound may be administered for at least 1 week, 2 weeks, 3 weeks, 4 weeks, 5
weeks, 6 weeks,
7 weeks or 8 weeks. In some embodiments, the method further comprises
detecting the steady
state plasma level of the substituted compound, or the level of the
substituted compounds within
red blood cell membrane to determine the incorporation level of the
substituted compound. In
some such embodiments, the plasma level of the substituted compound reaches a
steady state after
1, 2, 3 or 4 weeks. In one embodiment, the plasma level of the substituted
compound may reach a
steady state within 15 days, 20 days, 30 days, 40 days, 50 days or 60 days.
[0087] In some embodiments, the dosage of the substituted compound is in
the range
from about 10 mg/kg to about 200 mg/kg, or from about 20 mg/kg to about 100
mg/kg. In some
embodiments, the dosage of the substituted compound is in the range from about
30 mg/kg to
about 80 mg/kg. In some embodiments, the daily dose of the substituted
compound is in the range
of about 1 g to about 10 g. In some embodiments, the daily dose of the
substituted compound is
about 1.8g or about 9g. In some embodiments, the daily dose of the substituted
compound is about
1.8g. In one embodiment, the daily dose of the substituted compound is about
4.5g administered
twice a day. In another embodiment, the daily dose of the substituted compound
is about 2.7g
administered twice a day.
[0088] In some embodiments, the substituted compound is co-administered
to the
subject with at least one antioxidant. In some embodiments, the antioxidant is
selected from
Coenzyme Q, idebenone, mitoquinone, mitoquinol, plastoquinone, resveratrol,
vitamin E, and
vitamin C, and combinations theneof. In some such embodiments, the antioxidant
may be taken
concurrently, prior to, or subsequent to the administration of the substituted
compound. In some
embodiments, the antioxidant and the substituted compound may be in a single
dosage form. In
some embodiments, the single dosage form is selected from the group consisting
of a pill, a tablet,
and a capsule.
Substituted compounds
[0089] In some embodiments, the substituted compound comprises at least
one
isotope, and the amount of the isotope is significantly above the naturally-
occurring abundance
level of the isotope. For example, in some embodiments, the amount of the
isotope is two or more
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times greater than the naturally-occurring abundance level of the isotope. In
some such
embodiments, the substituted compound comprises an amount of deuterium that is
significantly
above the naturally-occurring abundance level of the deuterium. For example,
in some
embodiments, the amount of the deuterium in the substituted compound is two or
more times
greater than the naturally-occurring abundance level of the deuterium.
[0090] In some embodiments, the substituted compound is an isotopically
modified
polyunsaturated fatty acid ester. In some embodiments, the substituted
compound is an
isotopically modified polyunsaturated fatty acid. In some embodiments, the
polyunsaturated fatty
acid ester is a triglyceride, a diglyceride, a monoglyceride or an alkyl
ester. In some embodiments,
the polyunsaturated fatty acid ester is an ethyl ester.
[0091] The term "substituted compound" as used herein, refers to a
compound that is
modified by substitution at one or more positions to reduce the rate at which
the compound is
oxidized. The modification can be an isotopic substitution or a non-isotopic
chemical
modification. Isotopic substitution can refer to one or more substitutions
with an isotope such as
deuterium or 13C. Non-isotopic modification can refer to substitution at an
allylic hydrogen with
another chemical group or changing the position of an unsaturated bond to
eliminate an allylic
hydrogen position to reduce oxidation of the substituted compound.
[0092] The term "polyunsaturated lipid" as used herein, refers to a
lipid that contains
one or more unsaturated bonds, such as a double or a triple bond, in its
hydrophobic tail. The
polyunsaturated lipid may be a polyunsaturated fatty acid (PUFA) or ester
thereof.
[0093] The term "bis-allylic position" as used herein, refers to the
position of the
polyunsaturated lipid, such as polyunsaturated fatty acid or ester thereof,
that corresponds to the
methylene groups of 1,4-diene systems. Examples of polyunsaturated lipids
having deuterium at
one or more bis-allylic positions include but are not limited to 11,11-
Dideutero-cis,cis-9,12-
Octadecadienoic acid (11,11-Dideutero-(9Z,12Z)-9,12-Octadecadienoic acid; D2-
LA); and
11,11,14,14- Tetradeutero-ci s,ci s,cis-9,12,15-Octadecatrienoi c acid
(11,11,14,14-Tetradeutero-
(9Z,12Z,15Z)-9,12,15-Octadecatrienoic acid; D4-ALA).
[0094] The term "pro-bis-allylic position" as used herein, refers to the
methylene
group in a compound that becomes the bis-allylic position upon desaturation.
For example, some
sites which are not bis-allylic in precursor PUFAs become bis-allylic upon
biochemical
transformation. The pro-bis-allylic positions, in addition to being
deuterated, can be further
substituted by carbon-13, each at levels of isotopic abundance above the
naturally-occurring
abundance level. For example, the pro-bis-allylic positions, in addition to
existing bis-allylic
positions, can be reinforced by isotopic substitution as shown below in
Formula (1), wherein IV
is -OH, -0-alkyl, -amine, -S-alkyl, or ¨0-cation (e.g., cation being Na + or
IC1); m is 0 to 10; n is
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1 to 5; and p is 0 to 10. In Formula (1), the position of the X atom
represents the pro-bis-allylic
position, while the position of the Y atom represents the bis-allylic
position, each of XI, X2, Y',
and Y2 atoms may independently be hydrogen or deuterium atoms, and at least
one of X', X2, Y',
or Y2 atoms is deuterium. Each Y1 and Y2 for each n unit can independently be
hydrogen or
deuterium atoms, and each X1 and X2 for each m unit can independently be
hydrogen or deuterium
atoms.
Ki y2
Y n CH2R (1)
xl 2 P I
_m
= OH, alkyl, Salkyl, amine, or cation;
Cen101
each Y1 1 2
each X1 and X2 (f8r soek unit) rilg<arr'igfA4 or
[0095]
Another example of a substituted compound having bis-allylic and pro-bis-
allylic positions is shown in Formula (2), wherein any of the pairs of V-Yn
and/or XI-Xm
independently represent the bis-allylic and pro-bis-allylic positions of PUFAs
respectively and
these positions may contain deuterium atoms. X1, X2, ... X, Y', Y2, atoms
can
independently be hydrogen or deuterium atoms, and at least one or more of XI,
)(2, yl,
Y2, ...r atoms is deuterium. In some embodiments, at least one of Y1, Y2, ...
r atoms is
deuterium. In some embodiments, p is 0, 1 or 2. In some embodiments, m is 0,
1,2, or 3. In some
embodiments, n is 1, 2, 3, or 4. In some embodiments, n is greater than 1. In
some embodiments,
n is less than 4.
0
t-ic
/-\ /-y \11/ CH2
(2
i y )
y2 yn-lyn )(2 xrti- m
poi = OH, Oalkyl, amine, SH, Salkyl, or Ocation; Y1
toy
R = 1-1--1:?-13. w 67W X to X, = H or D; m =1-10; n=1-6; and p =0 -10
n
[0096] A
substituted compound as described herein can be a polyunsaturated lipid that
has at least one substitution that reduces oxidation of the substituted
compound. In some
embodiments, the substituted compound is isotopically modified to reduce
oxidation. In some
embodiments, the substituted compound is non-isotopically modified at one or
more positions to
reduce oxidation. The substituted compound, for example, isotopically modified
PUFAs such as
deuterated PUFAs may reduce oxidation by at least 5%, 10%, 15%, 20%, 25%, 30%,
35%, 40%,
45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90% or 95%.
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[0097] In some embodiments, the substituted compound as described herein
comprises
an isotopically modified polyunsaturated fatty acid, isotopically modified
polyunsaturated fatty
acid ester, isotopically modified polyunsaturated fatty acid thioester,
isotopically modified
polyunsaturated fatty acid amide, isotopically modified polyunsaturated fatty
acid mimetic, or
isotopically modified polyunsaturated fatty acid pro-drug. In some
embodiments, the substituted
compound as described herein can be an isotopically modified polyunsaturated
fatty acid or fatty
acid ester. In some embodiments, the substituted compound can be an
isotopically modified
naturally occurring PUFA. In some embodiments, the substituted compound can
have conjugated
double bonds. In some embodiments, the substituted compound is an isotopically
modified
polyunsaturated fatty acid. In some embodiments, the substituted compound is
an isotopically
modified polyunsaturated fatty acid thioester. In some embodiments, the
substituted compound is
an isotopically modified polyunsaturated fatty acid amide. In some
embodiments, the substituted
compound is an isotopically modified polyunsaturated fatty acid mimetic. In
some embodiments,
the substituted compound is an isotopically modified polyunsaturated fatty
acid prodrug.
[0098] In some embodiments, the substituted compound can be a deuterated
polyunsaturated lipid. In some embodiments, the substituted compound can be a
deuterated
polyunsaturated fatty acid, a deuterated polyunsaturated fatty acid ester, a
deuterated
polyunsaturated fatty acid thioester, a deuterated fatty acid amide, a
deuterated polyunsaturated
fatty acid mimetic, a deuterated polyunsaturated fatty acid pro-drug, or
combinations thereof.
[0099] In some embodiments, the substituted compound is deuterated at
one or more
bis-allylic positions. In some embodiments, the substituted compound is
further deuterated at one
or more pro-bis-allyl positions.
[0100] In some embodiments, the substituted compound is a co-3 fatty
acid, a co-6 fatty
acid, a co-3 fatty acid ester, a co-6 fatty acid ester, a co-3 fatty acid
amide, a o.)-6 fatty acid amide, a
co-3 fatty acid thioester, or a co-6 fatty acid thioester, or combinations
thereof. In some
embodiments, the substituted compound is a co-3 fatty acid, a (1)-3 fatty acid
ester, a co-3 fatty acid
amide, a co-3 fatty acid thioester, a prodrug thereof, or a combination
thereof. In some
embodiments, the substituted compound is a a)-6 fatty acid, a w-6 fatty acid
ester, a co-6 fatty acid
amide, a co-6 fatty acid thioester, a prodrug thereof, or combinations
thereof. In some
embodiments, the substituted compound is a linoleic acid, a linolenic acid, an
arachidonic acid,
an eicosapentaenoic acid, a docosahexaenoic acid, or an ester, amide,
thioester, or prodrug thereof,
or combinations thereof.
[0101] In some embodiments, the subject also ingests at least one of an
unsubstituted
polyunsaturated fatty acid and an unsubstituted polyunsaturated fatty acid
ester.
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[0102] In some embodiments, the amount of the substituted compound is
about 5% or
greater than the total amount of the polyunsaturated fatty acids and
polyunsaturated fatty acid
esters administered or delivered to the subject. In some embodiments, the
amount of the
substituted compound is about 10% or greater than the total amount of the
polyunsaturated fatty
acids and polyunsaturated fatty acid esters administered to the patient. In
some embodiments, the
amount of the substituted compound is about 15% or greater than the total
amount of the
polyunsaturated fatty acids and polyunsaturated fatty acid esters administered
to the subject. In
some other embodiments, the amount of the substituted compound is equal to or
less than about
1% of the total amount of the polyunsaturated fatty acids and polyunsaturated
fatty acid esters
administered or delivered to the subject.
[0103] In some embodiments, the polyunsaturated fatty acid,
polyunsaturated fatty
acid ester, polyunsaturated fatty acid thioester, polyunsaturated fatty acid
amide, polyunsaturated
fatty acid mimetic, or polyunsaturated fatty acid pro-drug can be a naturally
occurring PUFA. In
some embodiments, the polyunsaturated fatty acid, polyunsaturated fatty acid
ester,
polyunsaturated fatty acid thioester, polyunsaturated fatty acid amide,
polyunsaturated fatty acid
mimetic, or polyunsaturated fatty acid pro-drug can have conjugated double
bonds.
[0104] In some embodiments, the substituted compound is deuterated at
one or more
positions. In some embodiments, the substituted compound is deuterated at one
or more bis-allylic
positions. In some embodiments, the polyunsaturated fatty acid,
polyunsaturated fatty acid ester,
polyunsaturated fatty acid thioester, polyunsaturated fatty acid amide,
polyunsaturated fatty acid
mimetic, or polyunsaturated fatty acid pro-drug is deuterated at one or more
positions. In some
embodiments, the polyunsaturated fatty acid, polyunsaturated fatty acid ester,
polyunsaturated
fatty acid thioester, polyunsaturated fatty acid amide, polyunsaturated fatty
acid mimetic, or
polyunsaturated fatty acid pro-drug is deuterated at one or more bis-allylic
positions.
[0105] In some embodiments, the substituted compound is a fatty acid or
fatty acid
ester. In some such embodiments, the ester may be a triglyceride, a
diglyceride, a monog,lyceride,
or an alkyl ester. In some further embodiments, the polyunsaturated fatty acid
ester is a methyl or
ethyl ester.
[0106] In some embodiments, the deuterated fatty acid or fatty acid
ester are co-
administered to a patient with non-deuterated fatty acids or fatty acid
esters.
[0107] In some embodiments, the substituted compound comprises between
about
lwt% to about 100wt%, about 5wt% to about 90wt%, about lOwt% to about 50W%,
about 20wt%
to about 40wt% of the total amount of polyunsaturated fatty acid,
polyunsaturated fatty acid ester,
polyunsaturated fatty acid thioester, polyunsaturated fatty acid amide,
polyunsaturated fatty acid
mimetic, and polyunsaturated fatty acid pro-drug administered or delivered to
the patient. In some
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embodiments, the substituted compound comprises between about 10 wt% and about
40 wt% of
the total amount of polyunsaturated fatty acid, polyunsaturated fatty acid
ester, polyunsaturated
fatty acid thioester, polyunsaturated fatty acid amide, polyunsaturated fatty
acid mimetic, and
polyunsaturated fatty acid pro-drug administered to the patient. In some
embodiments, the
substituted compound comprises about 1 we/0, 2wt%, 3wt%, 4wt%, 5wt%, 6wt%,
7wt%, 8wt%,
9wt%, lOwt%, 15wt%, 20wt% or more of the total amount of polyunsaturated fatty
acid,
polyunsaturated fatty acid ester, polyunsaturated fatty acid thioester,
polyunsaturated fatty acid
amide, polyunsaturated fatty acid mimetic, and polyunsaturated fatty acid pro-
drug administered
or delivered to the patient. In some further embodiments, the substituted
compound is a deuterated
fatty acid or fatty acid ester.
[0108] In some embodiments, the deuterated fatty acid or fatty acid
ester comprises
between about lwt% to about 100wt%, about 5wt% to about 90wt%, about 1 Owt% to
about
50wt%, about 20wt% to about 40wt% of the total amount of fatty acids or fatty
acid esters
administered or delivered to the subject. In some embodiments, the deuterated
fatty acid or fatty
acid ester comprises about Iwt%, 2vvt%, 3wt%, 4wt%, 5wt%, 6wt%, 7vvt%, 8wt%,
9vvt%, lOwt%,
15wt%, 20wt% or more of the total amount of fatty acids or fatty acid esters
administered or
delivered to the subject.
[0109] In some embodiments, a cell or tissue of the patient maintains a
sufficient
concentration of the deuterated fatty acid or fatty acid ester to prevent or
reduce autoxidation of
the naturally occurring non-deuterated fatty acid or fatty acid ester.
[0110] In some embodiments, the deuterated substituted compound has an
isotopic
purity in the range of about 20% to about 99%.
101111 In some embodiments, the fatty acid or fatty acid ester is one or
more selected
from the group consisting of 11,11-D2-linolenic acid, 14,14-D2-linolenic acid,
11,11,14,14-D4-
linolenic acid, 11,11-D2-linoleic acid, 14,14-D2-linoleic acid, and
11,11,14,14-D4-linoleic acid.
In some embodiments, the substituted compound is an omega-3 fatty acid or an
omega-3 fatty
acid ester. In some embodiments, the substituted compound is an omega-6 fatty
acid or an omega-
6 fatty acid ester.
[0112] In some embodiments, the substituted compound is a linoleic acid,
a linolenic
acid, an arachidonic acid (ARA), a docosahexaenoic acid (DHA), or an
eicosapentaenoic acid,
(EPA), or combinations thereof. In some embodiments, the substituted compound
is an
arachidonic acid, a docosahexaenoic acid, an eicosapentaenoic acid containing
one or more
deuterium. In some embodiments, the substituted compound is an arachidonic
acid, a
docosahexaenoic acid, an eicosapentaenoic acid, each containing one or more
deuterium at one or
more bis-allylic positions.
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[0113] In
some embodiments, the substituted compound is selected from the group
consisting of 11,11-D2-linolenic acid, 14,14-D2-linolenic acid, 11,11,14,14-D4-
linolenic acid,
11,11-D2-linoleic acid, an ester thereof, and a combination thereof. In some
embodiments, the
substituted compound is selected from the group consisting of 7,7-D2-
arachidonic acid; 10,10-
D2-arachidonic acid; 13,13-D2-arachidonic acid; 7,7,10,10-D4-arachidonic acid;
7,7,13,13-D4-
arachidonic acid; 10,10,13,13-D4-arachidonic acid; 7,7,10,10,13,13-D6-
arachidonic acid;
7,7,10,10,13,13,16,16-D8-eicosapentaenoic acid;
6,6,9,9,12,12,15,15,18,18-D10-
docosahexaenoic acid; an ester thereof, and combinations thereof. In some
embodiments, the
substituted compound is 11,11-D2-linoleic acid ethyl ester. In some
embodiments, the substituted
compound is 11,11,14,14-D4-linolenic acid ethyl ester. In some embodiments,
the substituted
compound is 7,7,10,10,13,13-D6-arachidonic acid; 7,7,10,10,13,13,16,16-D8-
eicosapentaenoic
acid; 6,6,9,9,12,12,15,15,18,18-D10-docosahexaenoic acid; or ester thereof. In
some
embodiments, the substituted compound is 7,7,10,10,13,13-D6-arachidonic acid;
7,7,10,10,13,13,16,16-D8-eicosapentaenoic acid or ester thereof. In some
embodiments, the
substituted compound is 7,7,10,10,13,13,16,16-D8-eicosapentaenoic acid or
ester thereof. In some
embodiments, the substituted compound is 6,6,9,9,12,12,15,15,18,18-D10-
docosahexaenoic acid;
or ester thereof.
[0114] In
some embodiments, the fatty acid or fatty acid ester is an omega-3 fatty acid.
In some embodiments, the omega-3 fatty acid is alpha linolenic acid. In some
embodiments, the
omega-3 fatty acid is ARA. In some embodiments, the omega-3 fatty acid is EPA.
In some
embodiments, the omega-3 fatty acid is DHA.
[0115] In
some embodiments, the fatty acid or fatty acid ester is an omega-6 fatty acid.
In some embodiments, the omega-6 fatty acid is linoleic acid. In some
embodiments, the omega-
6 fatty acid is gamma linolenic acid, dihomo gamma linolenic acid, arachidonic
acid, or
docosatetraenoic acid. In some embodiments, the fatty acid or fatty acid ester
is an omega-6 ARA.
In some embodiments, the fatty acid or fatty acid ester is an omega-6 DHA. In
some
embodiments, the fatty acid or fatty acid ester is an omega-6 EPA.
[0116] The
substituted compound that is isotopically reinforced at oxidation sensitive
positions as described by way of the structures above are heavy isotope
enriched at said positions
as compared to the natural abundance of the appropriate isotope. In some
embodiments, the
substituted compound has the deuterium atom present at a level greater than
its natural abundance
level. Deuterium has a natural abundance of about 0.0156%. Thus, a substituted
compound having
greater than the natural abundance of deuterium has greater than 0.0156% of
its hydrogen atoms
replaced or "reinforced" with deuterium, such as 0.02%, but preferably about
5%, 10%, 15%,
20%, 25%, 30%, 35%, 40%, 45%, 50%, 65%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,
95%, or
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100% of deuterium with respect to one or more hydrogen atoms in each
substituted compound
molecule. In other embodiments, the percentage of total hydrogen atoms in a
substituted
compound that is reinforced with deuterium is at least 0.02%, 0.03% (about
twice natural
abundance), 0.05%, 0.1%, 1%, 2%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%,
50%, 65%,
60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%.
[0117] In some aspects, a composition of substituted compound contains
both
isotopically modified polyunsaturated lipid and isotopically unmodified
polyunsaturated lipid. In
some embodiments, isotopic purity refers to the percentage of molecules of an
isotopically
modified polyunsaturated lipid in the composition relative to the total number
of molecules of the
isotopically modified polyunsaturated lipid plus polyunsaturated lipid with no
heavy atoms. For
example, the isotopic purity may be about 5%, 10%, 15%, 20%, 25%, 30%, 35%,
40%, 45%,
50%, 65%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the molecules of
isotopically
modified polyunsaturated lipid relative to the total number of molecules of
both the isotopically
modified polyunsaturated lipid plus polyunsaturated lipid with no heavy atoms.
In other
embodiments, the isotopic purity is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,
40%, 45%,
50%, 65%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some
embodiments,
isotopic purity of the polyunsaturated lipid may be from about 10%-100%, 10%-
95%, 10%-90%,
10%-85%, 10%-80%, 10%-75%, 10%-70%, 10%-65%, 10%-60%, 10%-55%, 10%-50%, 10%-
45%, 10%-40%, 10%-35%, 10%-30%, 10%-25%, or 10%-20% of the total number of
molecules
of the polyunsaturated lipid in the composition. In other embodiments,
isotopic purity of the
polyunsaturated lipid may be from about 15%-100%, 15%-95%, 15%-90%, 15%-85%,
15%-80%,
15%-75%, 15%-70%, 15%-65%, 15%-60%, 15%-55%, 15%-50%, 15%-45%, 15%-40%, 15%-
35%, 15%-30%, 15%-25%, or 15%-20% of the total number of molecules of the
polyunsaturated
lipid in the composition. In some embodiments, isotopic purity of the
polyunsaturated lipid may
be from about 20%4 00%, 20%-95%, 20%-90%, 20%-85%, 20%-80%, 20%-75%, 20%-70%,
20%-65%, 20%-60%, 20%-55%, 20%-50%, 20%-45%, 20%-40%, 20%-35%, 20%-30%, or 20%-
25% of the total number of molecules of the polyunsaturated lipid in the
composition. Two
molecules of an isotopically modified polyunsaturated lipid out of a total of
100 total molecules
of isotopically modified polyunsaturated lipid plus polyunsaturated lipid with
no heavy atoms can
have 2% isotopic purity, regardless of the number of heavy atoms the two
isotopically modified
molecules contain.
[0118] In some aspects, an isotopically modified PUFA molecule may
contain one
deuterium atom, such as when one of the two hydrogens in a methylene group is
replaced by
deuterium, and thus may be referred to as a "DI" PUFA. Similarly, an
isotopically modified
PUFA molecule may contain two deuterium atoms, such as when the two hydrogens
in a
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methylene group are both replaced by deuterium, and thus may be referred to as
a "D2" PUFA.
Similarly, an isotopically modified PUFA molecule may contain three deuterium
atoms and may
be referred to as a "D3" PUFA. Similarly, an isotopically modified PUFA
molecule may contain
four deuterium atoms and may be referred to as a "D4" PUFA. In some
embodiments, an
isotopically modified PUFA molecule may contain five deuterium atoms or six
deuterium atoms
and may be referred to as a "D5" or "D6" PUFA, respectively.
[0119] The number of heavy atoms in a molecule, or the isotopic load,
may vary. For
example, a molecule with a relatively low isotopic load may contain about 1,
2, 3, 4, 5, or 6
deuterium atoms. A molecule with a moderate isotopic load may contain about
10, 11, 12, 13, 14,
15, 16, 17, 18, 19, or 20 deuterium atoms. In a molecule with a very high
load, every hydrogen
may be replaced with a deuterium. Thus, the isotopic load refers to the
percentage of heavy atoms
for that type of atom in each substituted compound or polyunsaturated lipid
molecule. For
example, the isotopic load may be about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%,
65%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the number of the same
type of
atoms in comparison to a substituted compound or polyunsaturated lipid with no
heavy atoms of
the same type (e.g. hydrogen would be the "same type" as deuterium). In some
embodiments, the
isotopic load is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,
65%, 60%, 65%,
70%, 75%, 80%, 85%, 90%, 95%, or 100%. Unintended side effects are expected to
be reduced
where there is high isotopic purity in a substituted compound or
polyunsaturated lipid composition
but low isotopic load in a given molecule. For example, the metabolic pathways
will likely be
less affected by using a substituted compound or polyunsaturated lipid
composition with high
isotopic purity but low isotopic load.
[0120] One will readily appreciate that when one of the two hydrogens of
a methylene
group is replaced with a deuterium atom, the resultant compound may possess a
stereo center. In
some embodiments, it may be desirable to use racemic compounds. In other
embodiments, it may
be desirable to use enantiomerically pure compounds. In additional
embodiments, it may be
desirable to use diastereomerically pure compounds. In some embodiments, it
may be desirable
to use mixtures of compounds having enantiomeric excesses and/or
diastereomeric excesses of
about 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 65%, 60%, 65%, 70%,
75%, 80%,
85%, 90%, 95%, or 100%. In other embodiments, the enantiomeric excesses and/or
diastereomeric excesses is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,
45%, 50%, 65%,
60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100%. In some embodiments, it may
be
preferable to utilize stereochemically pure enantiomers and/or diastereomers
of embodiments -
such as when contact with chiral molecules is being targeted for attenuating
oxidative damage.
However, in many circumstances, non-chiral molecules are being targeted for
attenuating
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oxidative damage. In such circumstances, embodiments may be utilized without
concern for their
stereochemical purity. Moreover, in some embodiments, mixtures of enantiomers
and
diastereomers may be used even when the compounds are targeting chiral
molecules for
attenuating oxidative damage.
[0121] In some aspects, an isotopically modified polyunsaturated lipid
imparts an
amount of heavy atoms in a particular tissue. Thus, in some aspects, the
amount of heavy
molecules will be a particular percentage of the same type of molecules in a
tissue. For example,
the number of heavy molecules may be about 1%400% of the total amount of the
same type of
molecules in a tissue. In some aspects, 10-50% of the molecules are
substituted with the same
type of heavy molecules.
[0122] In some embodiments, the polyunsaturated lipid is deuterated at
one or more
bis-allylic positions. One example of a polyunsaturated lipid is an essential
PUFAs that is
isotopically modified at bis-allylic positions as shown below in Formula (3),
whereas R1 is
-0-alkyl, -OH, amine, -SH, -S-alkyl, or -0-cation (e.g., cation is Na l" or
IC); m is 1 to 10; n is 1
to 5; R is H or alkyl (e.g., C3F17). The bis-allylic positions, in addition to
deuteration, can be
further reinforced by carbon-13, each at levels of isotope abundance above the
naturally-occurring
abundance level. At each bis-allylic position in Formula (3), one or both Y1,
Y2 atoms in each n
unit are independently deuterium atoms. In some embodiments, n is 1, 2, 3, or
4. In some
embodiments, m is 1, 2,3, or 4.
CH2 \¨ (3)
/ y2
YI rn R1
_n
-
101231 Exact structures of compounds illustrated above are shown below
that provide
for both isotope reinforced n-3 (omega-3) and n-6 (omega-6) essential
polyunsaturated fatty acids,
and the PUFAs made from them biochemically by desaturation/elongation. Any one
of these
compounds may be used to slow oxidation. In the following compounds, the PUFAs
are
isotopically reinforced at oxidation sensitive sites and/or sites that may
become oxidation sensitive
upon biochemical desaturation/elongation. le may be H, alkyl, or cation (e.g.,
No+ or K+); R2
may be H or D; * represents either 12C or 13C.
[0124] Deuterated linoleic acids may include:
D R2
D R2 D R2
0 0
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R2
npi n pp 1
... , ..D ' R.. ,2, . . = . , = µ ........D =
F,,,i% D__,....õ...y,...
D
R2 R2
, =
R2
1 R2
F/ OR I
D ''.= 0 -'.= ' .=' D--:"'ir"
0 R2
R2 0
= .
10125] The per-deuterated linoleic acid below may be produced by
microbiological
methods, for example by growing in media containing deuterium and/or carbon-
13.
DD ODD D
D36,!:/).D .D,,,,,D. D Dy(..,,._.õ
OR1
D D . ¨...; 1)-- D let
D---/:=:.
D DDIE)D ¨
D D D D
101261 Deuterated arachidonic acids may include:
00 R1
. 0 OW =
---
D R2 '' `=--;
= ,---*
R2 0 OR1
00 R1 * . R2
D'''=i'
---- R2 0
R2 R2
0OR1 ...,.. - õ... ,..7., .,.---C,...:::0R
121 1
R2
I:r ''. --'-
R- R2 D
= .
101271 The per-deuterated arachidonic acid below may be produced by
microbiological methods, such as by growing in media containing deuterium
and/or carbon-13.
D D
D D DDD D
D3 > = .' -14. .1D . 00W
= D D'"`%(1----
c)...4-1<..,/
D0 DO D D DDDD
D
101281 Deuterated linolenic acids may include:
0
0
J.---%-....."...,..,, OR1
D = R2 "'..sl, y(OR1 = ..."
D
R2
= =
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0 0
fOR1
R2
= R2 D = OR1
D R*2 R2
0
R2 R2
ji.NOR1
* *
D D D D
R2 R2 R2 R2
0 0
2
jOR1
D= R2 D D R2
=
D D
R2 R2 R2
0
R2 0
,JOR1 R2
OR1
D , D R2 D =
R-
, and
. OR
D
R-
101291 Per-deuterated linolenic acid below may be produced by
microbiological
methods, such as growing in media containing deuterium and/or carbon-13.
D D D nD o\
je¨ 4100 -%15<L*-/D l'OR1
* D D * Dõõ * D
D3C D
D DO D
D D
101301 Deuterated polyunsaturated fatty acid and ester may also include:
0
OH
0
DD DO DD DD DD 0
H , and ester thereof.
101311 In a further embodiment, oxidation-prone bis-allylic sites of
substituted
compounds as described herein (e.g., PUFAs) can be protected against hydrogen
abstraction by
moving bis-allylic hydrogen-activating double bonds further apart, thus
eliminating the bis-allylic
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positions while retaining certain PUFA functionality as shown below. These
PUFA mimetics have
no bis-allylic positions.
H3C 0, H3C
(.1-011
\
Octade124:dafrckc acid Octadeca-7õ1111;15ttitimilic acid
0
= H; alkyl; n = 1-4; = 1-12
IR 5 Hi 007; = H; alkyl; n =1-4; m = 1-12
[0132] in a further embodiment, oxidation-prone bis-allylic sites of
substituted
compounds as described herein (e.g., PUFAs) can be protected against hydrogen
abstraction by
incorporating heteroatoms with valence III (e.g., S, 0), thus eliminating the
bis-allylic hydrogens
as shown below. These PUFA mimetics also have no bis-allylic hydrogens.
H3C
/
X 0 10-P-4#0-1-onylswifanyl-deoriOz 4cid X E 10-(2-
But=14nylguifervi.vinylulfany4)4 4 tici
X 0; 10-Hopt-1-finvioxy-dtlfA-tmakada X E 0:10-(2-But-1-
onyioxpvinyloxy).dgQ4onok odd
bw
R
R = 03147. :1414 11.!, W-11
[0133] In a further embodiment, PUFA mimetics, i.e. compounds
structurally similar
to natural PUFAs but more resistant to oxidation because of the structural
differences, can be
employed for the above mentioned purposes. Oxidation-prone bis-allylic sites
of PUFAs can be
protected against hydrogen abstraction by di-methylation or halogenation as
shown below. The
hydrogen atoms on the methyl groups may optionally be halogens, such as
fluorine, or deuterium.
These PUFA mimetics are dim ethylated at bis-allylic sites.
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H3C
H3C
\ 3C CH / CH3
CH3 OH OH
¨ CH3
¨ CH3
11,11,14,14-Tetramethyl-
11,11-Dimethyl-octadeca-9,12-dienoic acid octadeca-9,12,15-trienoic acid
0
/
/ x2 CH214
R0
R1 HCH3CH2IT %Ri X1
-n
R -n
R = H or C R1 = H or alkyl; n = 1-5; m = 1-
12;
R = H or C3H7: R1 = H or alkyl; xi, x2 are Oldependently F, Cl., Br, or I
n = 1-5; m = 1-12
101341 In a further embodiment, oxidation-prone bis-allylic sites of
PUFAs can be
protected against hydrogen abstraction by alkylation as shown below. These PU
FA mimetics are
dialkylated at bis-allylic sites.
H3C HC
OHOH
10-(144504eirOodopttp*ftAkentaicaciii
10-{1-p-pailat-tteeryylowdtpum*
vimApqptsomp*dol-94moicedd
5r1-1C4/4 )Ri
ft
111 03117,
101351 In a further embodiment, cyclopropyl groups can be used instead
of double
bonds, thus rendering the acids certain functions while eliminating the bis-
allylic sites as shown
below. These PUFA mimetics have cyclopropyl groups instead of double bonds.
H3C
OH
H3C
8-(2-0-FitaWyclopropyimethyi)-
cydapropyikociancit odd
842-[2-(204.idopropy1mediy1)-
Ri dddritrodP*10004141401M140411damOt add
R CA. RI 210' : 1-5m=1-12
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[0136] In a further embodiment, 1,2-substituted cyclobutyl groups in
appropriate
conformations can be used instead of double bonds, thus rendering the acids
certain functions
while eliminating the bis-allylic sites as shown below. These PUFA mimetics
have 1,2-cyclobutyl
groups instead of double bonds.
OH
1.13C
8-1,24.2-Pagrwknotatioutprattro0- OH
cyclobutyli-octanciit acid
8-[2-R24V2-FfitlysNayscdidaiiWmaitty)))-
t R1
CHt=1:7 cyclobutylmethyl)-cyclobutylycittaliek acid
if!
R C3147, RI alkyl; n
.111; 1:11i
[0137] In a modification of the previous embodiment of mimetics with 1,2-
cyclobutyl
groups instead of double bonds, 1,3-substituted cyclobutyl groups in
appropriate conformations
can be used instead of double bonds, thus rendering the acids certain
functions while eliminating
the bis-allylic sites. The following PUFA mimetics have 1,3-cyclobutyl groups
instead of double
bonds.
HaCmyO
r_x_40
b"
8430ftnidovp.mhtioneittm)-
cyclobutyl]=octanoic acid i0P1
8-INPAPQRP~IMInitnititt61)-
cyclobutylmethy1]-cycktutylyocianciic acid
Ft9 R1
= H; alkyl; n = 1-5; m = 1-12
R CPT RI = H; alkyl; n = 1-5; m = 1-12
[0138] Bioisosteres are substituents or groups with similar physical or
chemical
properties which produce broadly similar biological properties to a chemical
compound. For
example, well known isosteres and/or bioisosteres for hydrogen include
halogens such as fluorine;
isosteres and/or bioisosteres of alkenes include alkynes, phenyl rings,
cyclopropyl rings,
cyclobutyl rings, cyclopentyl rings, cyclohexyl rings, thioethers, and the
like; isosteres and/or
bioisosteres of carbonyls include sulfoxides, sulfones, thiocarbonyls, and the
like; isosteres and/or
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bioisosteres of esters include amides, sulfonic acid esters, sulfonamides,
sulfinyl acid esters,
sulfinylamindes, and the like. Consequently, PUFA mimetics also include
compounds having
isosteric and/or bioisosteric functional groups.
[0139] In some embodiments, PUFAs and/or PUFA mimetics are formulated as
a pro-
drug for use in the various methods described herein. A pro-drug is a
pharmacological substance
that may itself have biological activity, but upon administration the pro-drug
is metabolized into
a form that also exerts biological activity. Many different types of pro-drugs
are known and they
can be classified into two major types based upon their cellular sites of
metabolism. Type I pro-
drugs are those that are metabolized intracellularly, while Type II are those
that are metabolized
extracellularly. It is well-known that carboxylic acids may be converted to
esters and various
other functional groups to enhance pharmacokinetics such as absorption,
distribution, metabolism,
and excretion. Esters are a well-known pro-drug form of carboxylic acids
formed by the
condensation of an alcohol (or its chemical equivalent) with a carboxylic acid
(or its chemical
equivalent). In some embodiments, alcohols (or their chemical equivalent) for
incorporation into
pro-drugs of PUFAs include pharmaceutically acceptable alcohols or chemicals
that upon
metabolism yield pharmaceutically acceptable alcohols. Such alcohols include,
but are not limited
to, propylene glycol, ethanol, isopropanol, 2-(2-ethoxyethoxy)ethanol
(Transcutol , Gattefosse,
Westwood, NJ. 07675), benzyl alcohol, glycerol, polyethylene glycol 200,
polyethylene glycol
300, or polyethylene glycol 400; polyoxyethylene castor oil derivatives (for
example,
polyoxyethyleneglyceroltriricinoleate or polyoxyl 35 castor oil (Cremophor EL,
BASF Corp.),
polyoxyethyleneglycerol oxystearate (Cremophor)RH 40 (polyethyleneglycol 40
hydrogenated
castor oil) or CremophoreRH 60 (polyethyleneglycol 60 hydrogenated castor
oil), BASF Corp.));
saturated polyglycolized glycerides (for example, Gelucire 35/10, Gelucire
44/14, Gelucire
46/07, Gelucire 50/13 or Gelucire 53/10, available from Gattefosse,
Westwood, N.J. 07675);
polyoxyethylene alkyl ethers (for example, cetomacrogol 1000); polyoxyethylene
stearates (for
example, PEG-6 stearate, PEG-8 stearate, polyoxyl 40 stearate NF, polyoxyethyl
50 stearate NF,
PEG-12 stearate, PEG-20 stearate, PEG-100 stearate, PEG-12 distearate, PEG-32
distearate, or
PEG-150 distearate); ethyl oleate, isopropyl palmitate, isopropyl myristate;
dimethyl isosorbide;
N-methylpyrrolidinone; paraffin; cholesterol; lecithin; suppository bases;
pharmaceutically
acceptable waxes (for example, camauba wax, yellow wax, white wax,
microcrystalline wax, or
emulsifying wax); pharmaceutically acceptable silicon fluids; soribitan fatty
acid esters (including
sorbitan laurate, sorbitan oleate, sorbitan palmitate, or sorbitan stearate);
pharmaceutically
acceptable saturated fats or pharmaceutically acceptable saturated oils (for
example, hydrogenated
castor oil (glyceryl-tris-12-hydroxystearate), cetyl esters wax (a mixture of
primarily C14-C18
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saturated esters of C14-C18 saturated fatty acids having a melting range of
about 43 -47 C), or
glyceryl monostearate) and combinations thereof.
[0140] In some embodiments, the fatty acid pro-drug is represented by
the ester P¨
B, wherein the radical P is a PUFA and the radical B is a biologically
acceptable molecule. Thus,
cleavage of the ester P¨B affords a PUFA and a biologically acceptable
molecule. Such cleavage
may be induced by acids, bases, oxidizing agents, and/or reducing agents.
Examples of
biologically acceptable molecules include, but are not limited to, nutritional
materials, peptides,
amino acids, proteins, carbohydrates (including monosaccharides,
disaccharides, polysaccharides,
glycosaminoglycans, and oligosaccharides), nucleotides, nucleosides, lipids
(including mono-, di-
and tri-substituted glycerols, glycerophospholipids, sphingolipids, and
steroids) and combinations
thereof.
[0141] In some embodiments, alcohols (or their chemical equivalent) for
incorporation
into pro-drugs of PUFAs include polyalcohols such as diols, triols, tetra-ols,
penta-ols, etc.
Examples of polyalcohols include ethylene glycol, propylene glycol, 1,3-
butylene glycol,
polyethylene glycol, methylpropanediol, ethoxydiglycol, hexylene glycol,
dipropylene glycol
glycerol, and carbohydrates. Esters formed from polyalcohols and PUFAs may be
mono-esters,
di-esters, tri-esters, etc. In some embodiments, multiply esterified
polyalcohols are esterified with
the same PUFAs. In other embodiments, multiply esterified polyalcohols are
esterified with
different PUFAs. In some embodiments, the different PUFAs are stabilized in
the same manner.
In other embodiments, the different PUFAs are stabilized in different manners
(such as deuterium
substitution in one PUFA and 13C substitution in another PUFA). In some
embodiments, one or
more PUFAs is an omega-3 fatty acid and one or more PUFAs is an omega-6 fatty
acid.
[0142] It is also contemplated that it may be useful to formulate PUFAs
and/or PUFA
mimetics and/or PUFA pro-drugs as salts for use, e.g., as pharmaceutically
acceptable salts. For
example, the use of salt formation as a means of tailoring the properties of
pharmaceutical
compounds is well known. See Stahl et al., Handbook of pharmaceutical salts:
Properties,
selection and use (2002) Weinheim/Zurich: Wiley-VCH/VHCA; Gould, Salt
selection for basic
drugs, hit. J. Pharm. (1986), 33:201-217. Salt formation can be used to
increase or decrease
solubility, to improve stability or toxicity, and to reduce hygroscopicity of
a drug product.
[0143] Formulation of PUFAs and/or PUFA esters and/or PUFA mirnetics
and/or
PUFA pro-drugs as salts can include any PUFA described herein.
[0144] It may be unnecessary to substitute all isotopically unmodified
PUFAs, such as
non-deuterated PUFAs, with isotopically modified PUFAs such as deuterated
PUFAs. In some
embodiments, is preferable to have sufficient isotopically modified PUFAs such
as D-PUFAs in
the membrane to prevent unmodified PUFAs such as H-PUFAs from sustaining a
chain reaction
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of self-oxidation. During self-oxidation, when one PUFA oxidizes, and there is
a non-oxidized
PUFA in the vicinity, the non-oxidized PUFA can get oxidized by the oxidized
PUFA. This may
also be referred to as autoxidation. In some instances, if there is a low
concentration, for example
"dilute" H-PUFAs in the membrane with D-PUFAs, this oxidation cycle may be
broken due to the
distance separating H-PUFAs. In some embodiments, the isotopically modified
PUFAs is present
in a sufficient amount to break an autoxidation chain reaction. To break the
autoxidation chain
reaction, for example, effective amounts of isotopically modified PUFAs may be
1-60%, 5-50%,
or 15-35% of the total molecules of the same type in the membrane.
Pharmaceutical Compositions
[0145] Some embodiments include pharmaceutical compositions comprising:
(a) a
safe and therapeutically effective amount of a substituted compound described
herein; and (b) a
pharmaceutically acceptable carrier, diluent, excipient or combination
thereof. In some
embodiments, the substituted compound is an isotopically modified
polyunsaturated acid (PUFA)
or an ester, thioester, amide, or other prodrug thereof, or combinations
thereof. In some further
embodiment, the isotopically modified PUFA is 11,11-D2-linoleic acid or an
ester thereof. In one
particular embodiment, the isotopically modified PUFA is 11,11-D2-linoleic
acid ethyl ester.
[0146] The compounds useful as described above can be formulated into
pharmaceutical compositions for use in treatment of various conditions or
disorders. Standard
pharmaceutical formulation techniques are used, such as those disclosed in
Remington's The
Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins
(2005), incorporated
by reference in its entirety.
101471 In addition to the selected compound useful as described above,
some
embodiments include compositions containing a pharmaceutically-acceptable
carrier. The term
"pharmaceutically-acceptable carrier", as used herein, means one or more
compatible solid or
liquid filler diluents or encapsulating substances, which are suitable for
administration to a
mammal. The term "compatible", as used herein, means that the components of
the composition
are capable of being commingled with the subject compound, and with each
other, in a manner
such that there is no interaction, which would substantially reduce the
pharmaceutical efficacy of
the composition under ordinary use situations. Pharmaceutically-acceptable
carriers must, of
course, be of sufficiently high purity and sufficiently low toxicity to render
them suitable for
administration preferably to an animal, preferably mammal being treated.
[0148] Pharmaceutically-acceptable carriers include, for example, solid
or liquid
fillers, diluents, hydrotropies, surface-active agents, and encapsulating
substances. Some
examples of substances, which can serve as pharmaceutically-acceptable
carriers or components
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thereof, are sugars, such as lactose, glucose and sucrose; starches, such as
corn starch and potato
starch; cellulose and its derivatives, such as sodium carboxymethyl cellulose,
ethyl cellulose, and
methyl cellulose; powdered tragacanth; malt; gelatin; talc; solid lubricants,
such as stearic acid
and magnesium stearate; calcium sulfate; vegetable oils, such as peanut oil,
cottonseed oil, sesame
oil, olive oil, corn oil and oil of theobroma; polyols such as propylene
glycol, glycerine, sorbitol,
mannitol, and polyethylene glycol; alginic acid; emulsifiers, such as the
TWEENS; wetting
agents, such sodium lauryl sulfate; coloring agents; flavoring agents;
tableting agents, stabilizers;
antioxidants; preservatives; pyrogen-free water; isotonic saline; and
phosphate buffer solutions.
[0149] Optional pharmaceutically-active materials may be included, which
do not
substantially interfere with the inhibitory activity of the compound. The
amount of carrier
employed in conjunction with the compound is sufficient to provide a practical
quantity of
material for administration per unit dose of the compound. Techniques and
compositions for
making dosage forms useful in the methods described herein are described in
the following
references, all incorporated by reference herein: Modem Pharmaceutics, 4th
Ed., Chapters 9 and
(Banker & Rhodes, editors, 2002); Lieberman et at., Pharmaceutical Dosage
Forms: Tablets
(1989); and Ansel, Introduction to Pharmaceutical Dosage Forms 8th Edition
(2004).
[0150] Various oral dosage forms can be used, including such solid forms
as tablets,
capsules, granules and bulk powders. Tablets can be compressed, tablet
triturates, enteric-coated,
sugar-coated, film-coated, or multiple-compressed, containing suitable
binders, lubricants,
diluents, disintegrating agents, coloring agents, flavoring agents, flow-
inducing agents, and
melting agents. Liquid oral dosage forms include aqueous solutions, emulsions,
suspensions,
solutions and/or suspensions reconstituted from non-effervescent granules, and
effervescent
preparations reconstituted from effervescent granules, containing suitable
solvents, preservatives,
emulsifying agents, suspending agents, diluents, sweeteners, melting agents,
coloring agents and
flavoring agents.
(0151] The pharmaceutically-acceptable carriers suitable for the
preparation of unit
dosage forms for peroral administration is well-known in the art. Tablets
typically comprise
conventional pharmaceutically-compatible adjuvants as inert diluents, such as
calcium carbonate,
sodium carbonate, mannitol, lactose and cellulose; binders such as starch,
gelatin and sucrose;
disintegrants such as starch, alginic acid and croscarmelose; lubricants such
as magnesium
stearate, stearic acid and talc. Glidants such as silicon dioxide can be used
to improve flow
characteristics of the powder mixture. Coloring agents, such as the FD&C dyes,
can be added for
appearance. Sweeteners and flavoring agents, such as aspartame, saccharin,
menthol, peppermint,
and fruit flavors, are useful adjuvants for chewable tablets. Capsules
typically comprise one or
more solid diluents disclosed above. The selection of carrier components
depends on secondary
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considerations like taste, cost, and shelf stability, which are not critical,
and can be readily made
by a person skilled in the art.
[0152] Per-oral compositions also include liquid solutions, emulsions,
suspensions,
and the like. The pharmaceutically-acceptable carriers suitable for
preparation of such
compositions are well known in the art. Typical components of carriers for
syrups, elixirs,
emulsions and suspensions include ethanol, glycerol, propylene glycol,
polyethylene glycol,
liquid sucrose, sorbitol and water. For a suspension, typical suspending
agents include methyl
cellulose, sodium carboxymethyl cellulose, AVICEL RC-591, tragacanth and
sodium alginate;
typical wetting agents include lecithin and polysorbate 80; and typical
preservatives include
methyl paraben and sodium benzoate. Peroral liquid compositions may also
contain one or more
components such as sweeteners, flavoring agents and colorants disclosed above.
[0153] Such compositions may also be coated by conventional methods,
typically with
pH or time-dependent coatings, such that the subject compound is released in
the gastrointestinal
tract in the vicinity of the desired topical application, or at various times
to extend the desired
action. Such dosage forms typically include, but are not limited to, one or
more of cellulose
acetate phthalate, polyvinylacetate phthalate, hydroxypropyl methyl cellulose
phthalate, ethyl
cellulose, Eudragit coatings, waxes and shellac.
[0154] Compositions described herein may optionally include other drug
actives or
supplements. For example, the pharmaceutical composition is administered
concomitantly with
one or more antioxidants. In some embodiments, the antioxidant is selected
from the group
consisting of Coenzyme Q, idebenone, mitoquinone, mitoquinol, vitamin E, and
vitamin C, and
combinations thereof. In some such embodiments, at least one antioxidant may
be taken
concurrently, prior to, or subsequent to the administration of the substituted
compound described
herein. In some embodiments, the antioxidant and the substituted compounds be
in a single dosage
form. In some embodiments, the single dosage form is selected from the group
consisting of a pill,
a tablet, and a capsule.
[0155] It will be understood by those of skill in the art that numerous
and various
modifications can be made without departing from the spirit of the present
disclosure. Therefore,
it should be clearly understood that the embodiments disclosed herein are
illustrative only and are
not intended to limit the scope of the present invention. Any reference
referred to herein is
incorporated by reference for the material discussed herein, and in its
entirety.
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EXAMPLES
Example 1
[0156] A study in the fatal neurodegenerative disease, Infantile
Neuroaxonal
Dystrophy (INAD) was initiated in March 2017. At study onset, the subject, a
2.5 year old female,
with a genetic mutation in both copies of the PLA2G6 gene, was mostly
unresponsive to stimuli
and unable to perform virtually any activity. All normal development
milestones previously
acquired had been lost. Prognosis was that a feeding tube would be required in
the near term
progression of the disease. The patient received dosing of a D-PUFA (11,11-D2-
linoleic acid), 1.8
g twice a day, for a six month trial under an Expanded Access (Compassionate
Use) protocol. As
she was unable to swallow a pill, soft gels containing the D-PUFA were placed
in warm liquid
(water or milk), pierced with a fork, and the active oil pressed into the
liquid, which was then
soaked into food or a cookie, and administered. Starting within a month of
dosing and continuing
until the current one year anniversary of dosing, the patient has improved.
FIG.! shows a detailed
list of development milestones lost by the subject prior to drug treatment,
and the observations of
the treating physicians versus the baseline assessment at the start of trial.
These results were
recorded in videotaped exams at baseline, 1 month, 3, 6, 9 and 12 months.
[0157] FIG. 1 summarizes the baseline and one year treatment status of
the patient
(degree of impairment: (0) for severely impaired, (+1) for moderately
impaired, (+2) for mildly
impaired or no impairment).
[0158] Within the first week, chronic constipation resolved. After 2
months on drug,
she regained sufficient bulbar function to terminate syringe feeding of
liquids and returned to
drinking from a child's sippy cup.
101591 After 4 months on drug, there were no drug related adverse events
of any sort
reported. One month and three month plasma level of deuterated polyunsaturated
fatty acid was
at steady state at approximately 44% total (dietary plus modified) linoleic
acid, indicating
excellent uptake and absorption of a therapeutic level of drug (>-10-15%).
Deuterated arachidonic
acid was observed as present in the plasma at 0.6% and 3.0% of total
arachidonic acid at month
one and three months; respectively. In addition, clinical examination at 3
months showed no
progression of disease since the start of the trial. Importantly, both
clinical examination video,
medical notes, and caregiver reports revealed the following observations:
11,11-D2-linoleic acid
was safe and well-tolerated at high dose in a 2 year old; the subject's
disease did not progress
(n=1); parents noted improvement as follows at 4 months including regained
ability to grasp spoon
and hold; regained ability to swallow using sipper cup (vs. syringe feeding);
can eat (chew and
swallow) food such as banana; salivation down 99%, essentially back to normal;
better muscle
strength (grasping and lifting rattle); newly responsive to verbal requests
during therapy; and
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constipation resolved, subject completely regular (constipation is a symptom
of PLA2G6
diseases).
[0160] After 6 months,
she improved in qualitative measures captured in videotaped
therapy sessions or exams, including eye tracking, responsiveness to verbal
commands, head
control, lifting, and reaching and grasping for her spoon (a lost skill). INAD
is a strictly
progressive disease, only worsening with time. Stabilization of regressions
and recovery of lost
milestones indicated effectiveness of the test treatment in a single, severely
affected INAD patient.
[0161] Stabilization
of progression of lost development milestones is a major advance
in therapy for INAD. In view of the clinical study results, these clear
reversals indicate that
substituted compounds as described herein are effective for treating a subject
having, or at risk
for, a disease or condition associated with an impaired Phospholipase A2 Group
VI (PLA2G6)
activity, and particularly the stabilized PUFA (11,11-D2-linoleic acid), in
patients with classical
WAD.
Example 2
[0162] In this
example, a case study of using 11,11-D2-linoleic acid ethyl ester to treat
a single patient with late onset Tay-Sachs disease (LOTS) and the study
results was reported.
[0163] Design/Methods:
11,11-D2-linoleic acid ethyl ester was administered to the
patient at 2.7 g (BID) and periodic repeat assessments including baseline
measurement of PK,
activities of daily living (ADL), 25 foot walk time (25FWT), and 6 minute walk
distance (6MWD)
were made. In particular, ADLs were measured individually on a scale of 0-5
across a 12 element
panel representing speech, strength, coordination, etc.
[0164] Results: 11,11-
D2-linoleic acid (D-LA) was elongated to 13,13-D2-
arachidonic acid (D-AA) and both deutenited PUFAs achieve significant plasma
levels and red
blood cell (RBC) membrane incorporation within 1 month of administration.
Improvements in
ADL, 25FWT, and 6MWD have also been seen. No major toxicities have been seen
(Table 1).
Table 1.
Time D-LA (irs) Composite ADL Score *of ADLs improved SPAWD
26,FINT
0-LA +1-1-LA 0411) (Sec)
Plasma RBC
Baseiine 0 0 0 112 12.7
30=d 36 28 +6 6 12.1
120 d nia nia +9 8 129 11.2
[0165] Conclusion:
Early efficacy signs of inhibition of disease progression and some
regression were seen. Furthermore, 11,11-D2-linoleic acid ethyl ester was well
tolerated with no
toxicities reported.
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